"

From 2012.igem.org

Revision as of 11:38, 15 September 2012 (view source) Asafbl (Talk | contribs) ← Older edit		Latest revision as of 11:17, 26 September 2012 (view source) Rachellev (Talk | contribs)
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	==Inbal==		==Inbal==
		+
		+	- Digest pTetO+m-cherry with XbaI+SpeI.<br>
		+	-6 minipreps for T7* RNAP+ter+pSB1AK3: the concentrations are: 71,139,126,97,40.5 and 54 ng/ul.<br>
		+	-Digest T7* RNAP+ter+pSB1AK3 with XbaI (x6).lycerol stock for this part. I also did a g<br>
		+	-colony PCR for T7+ter+pSB1AK3 (2700bp) with 2 suitable primers (total of 6 colony PCR reactions)- after the PCR, I ran the products on gel, and 1 band was positive! I have a positive clone! yay! now I can continue with the cloning!<br>
		+	-I digested T3 native RNAP with SpeI and XbaI and cleaned it by kit, the concentration was 31.4ng/ul.<br>
		+	- I also cleaned the PCR pTetO+m-cherry product by kit and the concentration was 20.6 ng/ul.<br>
		+	-I cleaned the products of yesterday restriction (of T7+ter+pSB1AK3) and the concentrations were:11.8,10.8,6,12,12.2 and 17.8 ng/ul.<br>
		+	- Also, today I ligated between T3 RNAP and pSB1AK3 plasmid (1 hour, at 37C), transformed it into TOP10 bacteria and plated 100ul and rest.<br>
		+	-I did a restriction reaction of T7* RNAP+ter+pSB1AK3 with PstI enzyme (15 minutes, 37C),for verifying what I have. I ran the products on a gel, and compared it to an uncut plasmid (approximately 3200bp on the gel), and the experimental sample had a band in 6kb approximately. This is a good result!<br>
		+	-3 starters of T7*+ter+pSB1AK3 (AMP resistance).<br>
		+	-GOOD NIGHT!
	==Asaf==		==Asaf==
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	==Hila==		==Hila==
		+	- Colony PCR for five positive colonies from each fragment transformation. <br>
		+	- I found two positive colonies for fragment 2, one positive colony for fragment 3, and three positive colonies for fragment 8. <br>
		+	- In addition I started to create the Gibson assembly fragments. I started with F4 and F5. F4 was amplified using two different AS primers: one with 100bp overlap and one with 400bp overlap. <br>
		+	- Overnight ligation to fragments 1C and 6C in pSB1C3_MCS.<br>
	==Lior==		==Lior==
-		+	We decided to check the T7 promoter parts with bacterias with PET system. we transformed the T7 promoter to the BL21 bacteria.<br>
		+	We also made colony PCR for our Positive control plasmids (pTetO with ALP/xyIE)
		+
	==Noa==		==Noa==
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	==Shahar==		==Shahar==
		+	-Purification PCR product of N4 RNAP (the one with pSB1A2 primers) by PCR cleaning kit.
		+	-Digest of N4 RNAP with XbaI and SpeI for creating sticky ends.
		+	-Clean the N4 RNAP after digestion by cleaning kit.
		+	-This insert (N4 RNAP) is ready for cloning.
	==Rachel==		==Rachel==
		+	- Digestion Bba_0015 with HindIII-HF & BamHI-HF to creat sticky ends.<br>
		+	- clean the products after restriction. the concentration was: 19.8 ng/ul
		+
		+
	}}		}}

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Latest revision as of 11:17, 26 September 2012

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Team:Technion/10_September_2012

March M T W T F S S       [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_March_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_March_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_March_2012&preload=Team:Technion/Template:NewDate&action=edit 3] [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_March_2012&preload=Team:Technion/Template:NewDate&action=edit 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_March_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_March_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_March_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_March_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_March_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_March_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_March_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_March_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_March_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_March_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_March_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_March_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_March_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_March_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_March_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_March_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/Team:Technion/21_March_2012 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_March_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_March_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_March_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_March_2012&preload=Team:Technion/Template:NewDate&action=edit 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_March_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_March_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_March_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_March_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_March_2012&preload=Team:Technion/Template:NewDate&action=edit 30] [http://2012.igem.org/wiki/index.php?title=Team:Technion/31_March_2012&preload=Team:Technion/Template:NewDate&action=edit 31]

April M T W T F S S             [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_April_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_April_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/Team:Technion/3_April_2012 3] [http://2012.igem.org/Team:Technion/4_April_2012 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_April_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_April_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_April_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_April_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_April_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_April_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_April_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_April_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_April_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/Team:Technion/14_April_2012 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_April_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_April_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_April_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_April_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_April_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_April_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_April_2012&preload=Team:Technion/Template:NewDate&action=edit 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_April_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/Team:Technion/23_April_2012 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_April_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/Team:Technion/25_April_2012 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_April_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_April_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/Team:Technion/28_April_2012 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_April_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_April_2012&preload=Team:Technion/Template:NewDate&action=edit 30]

May M T W T F S S   [http://2012.igem.org/Team:Technion/1_May_2012 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_May_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_May_2012&preload=Team:Technion/Template:NewDate&action=edit 3] [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_May_2012&preload=Team:Technion/Template:NewDate&action=edit 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_May_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_May_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_May_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_May_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_May_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/Team:Technion/10_May_2012 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_May_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_May_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_May_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_May_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_May_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_May_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_May_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_May_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_May_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_May_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/Team:Technion/21_May_2012 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_May_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_May_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_May_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_May_2012&preload=Team:Technion/Template:NewDate&action=edit 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_May_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_May_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_May_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_May_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_May_2012&preload=Team:Technion/Template:NewDate&action=edit 30] [http://2012.igem.org/wiki/index.php?title=Team:Technion/31_May_2012&preload=Team:Technion/Template:NewDate&action=edit 31]

June M T W T F S S         [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_June_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_June_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/Team:Technion/3_June_2012 3] [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_June_2012&preload=Team:Technion/Template:NewDate&action=edit 4] [http://2012.igem.org/Team:Technion/5_June_2012 5] [http://2012.igem.org/Team:Technion/6_June_2012 6] [http://2012.igem.org/Team:Technion/7_June_2012 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_June_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_June_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/Team:Technion/10_June_2012 10] [http://2012.igem.org/Team:Technion/11_June_2012 11] [http://2012.igem.org/Team:Technion/12_June_2012 12] [http://2012.igem.org/Team:Technion/13_June_2012 13] [http://2012.igem.org/Team:Technion/14_June_2012 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_June_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_June_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/Team:Technion/17_June_2012 17] [http://2012.igem.org/Team:Technion/18_June_2012 18] [http://2012.igem.org/Team:Technion/19_June_2012 19] [http://2012.igem.org/Team:Technion/20_June_2012 20] [http://2012.igem.org/Team:Technion/21_June_2012 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_June_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_June_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/Team:Technion/24_June_2012 24] [http://2012.igem.org/Team:Technion/25_June_2012 25] [http://2012.igem.org/Team:Technion/26_June_2012 26] [http://2012.igem.org/Team:Technion/27_June_2012 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_June_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_June_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_June_2012&preload=Team:Technion/Template:NewDate&action=edit 30]

July M T W T F S S             [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_July_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/Team:Technion/2_July_2012 2] [http://2012.igem.org/Team:Technion/3_July_2012 3] [http://2012.igem.org/Team:Technion/4_July_2012 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_July_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_July_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_July_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/Team:Technion/8_July_2012 8] [http://2012.igem.org/Team:Technion/9_July_2012 9] [http://2012.igem.org/Team:Technion/10_July_2012 10] [http://2012.igem.org/Team:Technion/11_July_2012 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_July_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_July_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_July_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_July_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_July_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/Team:Technion/17_July_2012 17] [http://2012.igem.org/Team:Technion/18_July_2012 18] [http://2012.igem.org/Team:Technion/19_July_2012 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_July_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_July_2012&preload=Team:Technion/Template:NewDate&action=edit 21] [http://2012.igem.org/Team:Technion/22_July_2012 22] [http://2012.igem.org/Team:Technion/23_July_2012 23] [http://2012.igem.org/Team:Technion/24_July_2012 24] [http://2012.igem.org/Team:Technion/25_July_2012 25] [http://2012.igem.org/Team:Technion/26_July_2012 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_July_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_July_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/Team:Technion/29_July_2012 29] [http://2012.igem.org/Team:Technion/30_July_2012 30] [http://2012.igem.org/Team:Technion/31_July_2012 31]

August M T W T F S S     [http://2012.igem.org/Team:Technion/1_August_2012 1] [http://2012.igem.org/Team:Technion/2_August_2012 2] [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_August_2012&preload=Team:Technion/Template:NewDate&action=edit 3] [http://2012.igem.org/Team:Technion/4_August_2012 4] [http://2012.igem.org/Team:Technion/5_August_2012 5] [http://2012.igem.org/Team:Technion/6_August_2012 6] [http://2012.igem.org/Team:Technion/7_August_2012 7] [http://2012.igem.org/Team:Technion/8_August_2012 8] [http://2012.igem.org/Team:Technion/9_August_2012 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_August_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_August_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/Team:Technion/12_August_2012 12] [http://2012.igem.org/Team:Technion/13_August_2012 13] [http://2012.igem.org/Team:Technion/14_August_2012 14] [http://2012.igem.org/Team:Technion/15_August_2012 15] [http://2012.igem.org/Team:Technion/16_August_2012 16] [http://2012.igem.org/Team:Technion/17_August_2012 17] [http://2012.igem.org/Team:Technion/18_August_2012 18] [http://2012.igem.org/Team:Technion/19_August_2012 19] [http://2012.igem.org/Team:Technion/20_August_2012 20] [http://2012.igem.org/Team:Technion/21_August_2012 21] [http://2012.igem.org/Team:Technion/22_August_2012 22] [http://2012.igem.org/Team:Technion/23_August_2012 23] [http://2012.igem.org/Team:Technion/24_August_2012 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_August_2012&preload=Team:Technion/Template:NewDate&action=edit 25] [http://2012.igem.org/Team:Technion/26_August_2012 26] [http://2012.igem.org/Team:Technion/27_August_2012 27] [http://2012.igem.org/Team:Technion/28_August_2012 28] [http://2012.igem.org/Team:Technion/29_August_2012 29] [http://2012.igem.org/Team:Technion/30_August_2012 30] [http://2012.igem.org/Team:Technion/31_August_2012 31]

September M T W T F S S           [http://2012.igem.org/Team:Technion/1_September_2012 1] [http://2012.igem.org/Team:Technion/2_September_2012 2] [http://2012.igem.org/Team:Technion/3_September_2012 3] [http://2012.igem.org/Team:Technion/4_September_2012 4] [http://2012.igem.org/Team:Technion/5_September_2012 5] [http://2012.igem.org/Team:Technion/6_September_2012 6] [http://2012.igem.org/Team:Technion/7_September_2012 7] [http://2012.igem.org/Team:Technion/8_September_2012 8] [http://2012.igem.org/Team:Technion/9_September_2012 9] [http://2012.igem.org/Team:Technion/10_September_2012 10] [http://2012.igem.org/Team:Technion/11_September_2012 11] [http://2012.igem.org/Team:Technion/12_September_2012 12] [http://2012.igem.org/Team:Technion/13_September_2012 13] [http://2012.igem.org/Team:Technion/14_September_2012 14] [http://2012.igem.org/Team:Technion/15_September_2012 15] [http://2012.igem.org/Team:Technion/16_September_2012 16] [http://2012.igem.org/Team:Technion/17_September_2012 17] [http://2012.igem.org/Team:Technion/18_September_2012 18] [http://2012.igem.org/Team:Technion/19_September_2012 19] [http://2012.igem.org/Team:Technion/20_September_2012 20] [http://2012.igem.org/Team:Technion/21_September_2012 21] [http://2012.igem.org/Team:Technion/22_September_2012 22] [http://2012.igem.org/Team:Technion/23_September_2012 23] [http://2012.igem.org/Team:Technion/24_September_2012 24] [http://2012.igem.org/Team:Technion/25_September_2012 25] [http://2012.igem.org/Team:Technion/26_September_2012 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_September_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_September_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_September_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_September_2012&preload=Team:Technion/Template:NewDate&action=edit 30]

October M T W T F S S [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_October_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_October_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_October_2012&preload=Team:Technion/Template:NewDate&action=edit 3] [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_October_2012&preload=Team:Technion/Template:NewDate&action=edit 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_October_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_October_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_October_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_October_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_October_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_October_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_October_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_October_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_October_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_October_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_October_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_October_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_October_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_October_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_October_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_October_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_October_2012&preload=Team:Technion/Template:NewDate&action=edit 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_October_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_October_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_October_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_October_2012&preload=Team:Technion/Template:NewDate&action=edit 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_October_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_October_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_October_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_October_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_October_2012&preload=Team:Technion/Template:NewDate&action=edit 30] [http://2012.igem.org/wiki/index.php?title=Team:Technion/31_October_2012&preload=Team:Technion/Template:NewDate&action=edit 31]

November M T W T F S S       [http://2012.igem.org/wiki/index.php?title=Team:Technion/1_November_2012&preload=Team:Technion/Template:NewDate&action=edit 1] [http://2012.igem.org/wiki/index.php?title=Team:Technion/2_November_2012&preload=Team:Technion/Template:NewDate&action=edit 2] [http://2012.igem.org/wiki/index.php?title=Team:Technion/3_November_2012&preload=Team:Technion/Template:NewDate&action=edit 3] [http://2012.igem.org/wiki/index.php?title=Team:Technion/4_November_2012&preload=Team:Technion/Template:NewDate&action=edit 4] [http://2012.igem.org/wiki/index.php?title=Team:Technion/5_November_2012&preload=Team:Technion/Template:NewDate&action=edit 5] [http://2012.igem.org/wiki/index.php?title=Team:Technion/6_November_2012&preload=Team:Technion/Template:NewDate&action=edit 6] [http://2012.igem.org/wiki/index.php?title=Team:Technion/7_November_2012&preload=Team:Technion/Template:NewDate&action=edit 7] [http://2012.igem.org/wiki/index.php?title=Team:Technion/8_November_2012&preload=Team:Technion/Template:NewDate&action=edit 8] [http://2012.igem.org/wiki/index.php?title=Team:Technion/9_November_2012&preload=Team:Technion/Template:NewDate&action=edit 9] [http://2012.igem.org/wiki/index.php?title=Team:Technion/10_November_2012&preload=Team:Technion/Template:NewDate&action=edit 10] [http://2012.igem.org/wiki/index.php?title=Team:Technion/11_November_2012&preload=Team:Technion/Template:NewDate&action=edit 11] [http://2012.igem.org/wiki/index.php?title=Team:Technion/12_November_2012&preload=Team:Technion/Template:NewDate&action=edit 12] [http://2012.igem.org/wiki/index.php?title=Team:Technion/13_November_2012&preload=Team:Technion/Template:NewDate&action=edit 13] [http://2012.igem.org/wiki/index.php?title=Team:Technion/14_November_2012&preload=Team:Technion/Template:NewDate&action=edit 14] [http://2012.igem.org/wiki/index.php?title=Team:Technion/15_November_2012&preload=Team:Technion/Template:NewDate&action=edit 15] [http://2012.igem.org/wiki/index.php?title=Team:Technion/16_November_2012&preload=Team:Technion/Template:NewDate&action=edit 16] [http://2012.igem.org/wiki/index.php?title=Team:Technion/17_November_2012&preload=Team:Technion/Template:NewDate&action=edit 17] [http://2012.igem.org/wiki/index.php?title=Team:Technion/18_November_2012&preload=Team:Technion/Template:NewDate&action=edit 18] [http://2012.igem.org/wiki/index.php?title=Team:Technion/19_November_2012&preload=Team:Technion/Template:NewDate&action=edit 19] [http://2012.igem.org/wiki/index.php?title=Team:Technion/20_November_2012&preload=Team:Technion/Template:NewDate&action=edit 20] [http://2012.igem.org/wiki/index.php?title=Team:Technion/21_November_2012&preload=Team:Technion/Template:NewDate&action=edit 21] [http://2012.igem.org/wiki/index.php?title=Team:Technion/22_November_2012&preload=Team:Technion/Template:NewDate&action=edit 22] [http://2012.igem.org/wiki/index.php?title=Team:Technion/23_November_2012&preload=Team:Technion/Template:NewDate&action=edit 23] [http://2012.igem.org/wiki/index.php?title=Team:Technion/24_November_2012&preload=Team:Technion/Template:NewDate&action=edit 24] [http://2012.igem.org/wiki/index.php?title=Team:Technion/25_November_2012&preload=Team:Technion/Template:NewDate&action=edit 25] [http://2012.igem.org/wiki/index.php?title=Team:Technion/26_November_2012&preload=Team:Technion/Template:NewDate&action=edit 26] [http://2012.igem.org/wiki/index.php?title=Team:Technion/27_November_2012&preload=Team:Technion/Template:NewDate&action=edit 27] [http://2012.igem.org/wiki/index.php?title=Team:Technion/28_November_2012&preload=Team:Technion/Template:NewDate&action=edit 28] [http://2012.igem.org/wiki/index.php?title=Team:Technion/29_November_2012&preload=Team:Technion/Template:NewDate&action=edit 29] [http://2012.igem.org/wiki/index.php?title=Team:Technion/30_November_2012&preload=Team:Technion/Template:NewDate&action=edit 30]

Ilya

- Did colony PCR for the pCP+RS+mCherry and pCP+RBS+mCherry clones that Rachel prepared for me. There were some positive results but then I realized that they are useless. The previous insert in the plasmid was also mCherry, so I had no way of telling the difference between the background clones and my clones judging by colony PCR.
- Put starters of the colonies I picked for the colony PCR in order to do a test with BamHI digestion tomorrow to find the correct clones.
- Planned some aspects of the gibson attempts that we are going to do this week with Roee and Hila. Tomorrow we will try the assembly of fragments 4 and 5 with 100bp and 400 bp overlaps with different incubation times in 37 degrees followed by the incubation in 50 degrees which is instructed in the gibson protocol.
- Prepared a 0.6% gel for testing the gel electrophoresis conditions for the gibson results.

Inbal

- Digest pTetO+m-cherry with XbaI+SpeI.
-6 minipreps for T7* RNAP+ter+pSB1AK3: the concentrations are: 71,139,126,97,40.5 and 54 ng/ul.
-Digest T7* RNAP+ter+pSB1AK3 with XbaI (x6).lycerol stock for this part. I also did a g
-colony PCR for T7+ter+pSB1AK3 (2700bp) with 2 suitable primers (total of 6 colony PCR reactions)- after the PCR, I ran the products on gel, and 1 band was positive! I have a positive clone! yay! now I can continue with the cloning!
-I digested T3 native RNAP with SpeI and XbaI and cleaned it by kit, the concentration was 31.4ng/ul.
- I also cleaned the PCR pTetO+m-cherry product by kit and the concentration was 20.6 ng/ul.
-I cleaned the products of yesterday restriction (of T7+ter+pSB1AK3) and the concentrations were:11.8,10.8,6,12,12.2 and 17.8 ng/ul.
- Also, today I ligated between T3 RNAP and pSB1AK3 plasmid (1 hour, at 37C), transformed it into TOP10 bacteria and plated 100ul and rest.
-I did a restriction reaction of T7* RNAP+ter+pSB1AK3 with PstI enzyme (15 minutes, 37C),for verifying what I have. I ran the products on a gel, and compared it to an uncut plasmid (approximately 3200bp on the gel), and the experimental sample had a band in 6kb approximately. This is a good result!
-3 starters of T7*+ter+pSB1AK3 (AMP resistance).
-GOOD NIGHT!

Asaf

I ran the fusion PCR of the Riboswitch with the different polymerases on a gel. This time I got the required bands (3.2 kb)
and also 2.7kb and 500 bp bands.

I ran the 56C Riboswitch+different polymerase on an extraction gel. I ran T7 of both 55 and 56C because I wasn't sure if he had a 3.2 kb band.

Hila

- Colony PCR for five positive colonies from each fragment transformation.
- I found two positive colonies for fragment 2, one positive colony for fragment 3, and three positive colonies for fragment 8.
- In addition I started to create the Gibson assembly fragments. I started with F4 and F5. F4 was amplified using two different AS primers: one with 100bp overlap and one with 400bp overlap.
- Overnight ligation to fragments 1C and 6C in pSB1C3_MCS.

Lior

We decided to check the T7 promoter parts with bacterias with PET system. we transformed the T7 promoter to the BL21 bacteria.
We also made colony PCR for our Positive control plasmids (pTetO with ALP/xyIE)

Noa

Evgeni

- Due to continuous failure of Bba_015 insertion in pPROLar plasmid I'll try new strategy: instead of double digestion of pPROLar I'll do sequential digestion: HindIII-HF restriction -> Heat inactivation of HindIII-HF at 80C -> BamHI-HF restriction.

- In addition I'll do new Bba_015 restriction with BamHI+HindIII, maybe ligation problems were due to problem with biobrick restriction (Done by Rachel)

- Overnight ligation of newly prepared pPROLar and Bba_015

Shahar

-Purification PCR product of N4 RNAP (the one with pSB1A2 primers) by PCR cleaning kit. -Digest of N4 RNAP with XbaI and SpeI for creating sticky ends. -Clean the N4 RNAP after digestion by cleaning kit. -This insert (N4 RNAP) is ready for cloning.

Rachel

- Digestion Bba_0015 with HindIII-HF & BamHI-HF to creat sticky ends.
- clean the products after restriction. the concentration was: 19.8 ng/ul