Team:Technion/3 September 2012

From 2012.igem.org

(Difference between revisions)

Latest revision as of 08:51, 20 September 2012

{{:Team:Technion/Template:Notebook|content=

Ilya

- Miniprep of the low copy number pCP plasmid.
- PCR of Fus2 and Fus2del from pSB3C5.

Inbal

- Mini-prep for the 3 starters I put yesterday. The concentrations are: 115, 197 and 150 ng/ul! :] (at least I know how to mini prep like a pro!)
- 4 starters of BBa_B0015 ( the insert is double terminators, the size is 2200bp roughly).
- Cleaning from gel the T7* RNAP (the one with pSB1A2 primers), the concentrations were l-o-w: about 7.5ng/ul after calculating it from just looking at the band's intensity (cause the NanoDrop wasn't accurate).
- clean the pSB1C3 plasmid after digestion with XbaI and PstI by cleaning kit.

Asaf

I cleaned the PCR products of the polymerase genes from the gel.

Hila

PCR reaction for fragments 1, 1C, 6, 6C using T = 55, 58, 60.
Cross your fingers…

Restriction with PacI and SpeI for fragments 2C, 3C, 4C, 5C, 7C, 8C and the confirmed MCS in the pSB1C3+MCS.

Lior

Noa

Evgeni

- No growth of starters again!!! Probably I'm doing some stupid mistake during their preparation, Orna will watch over me today during starters' preparation.

Shahar

-Clean the N$ & T3 RNAP after digestion by cleaning kit.
-Those inserts (N4 & T3 RNAP) are ready for cloning into pPROLAR plasmid.

Rachel

-Clean the PCR product of K1F RNAP (the one with pPROLAR primers)from the gel. The concentration was: 17 ng/ul.
-Digest of K1F RNAP with HindIII and NHeI for creating sticky ends.
-Clean the K1F RNAP after digestio by cleaning kit. The final concentration: 25 ng/ul
-This insert (K1F RNAP) is ready for cloning into pPROLAR plasmid.

@@ Line 10: / Line 10: @@
 - Cleaning from gel the T7* RNAP (the one with pSB1A2 primers), the concentrations were l-o-w: about 7.5ng/ul after calculating it from just looking at the band's intensity (cause the NanoDrop wasn't accurate). <br>
 - clean the pSB1C3 plasmid after digestion with XbaI and PstI by cleaning kit. <br>
--mini prep for the BBa_B0015 biobrick , the concentrations are: 192.5, 252.5, 143.5 and 179 ng/ul (GREAT!).<br>
--Digestion of BBa_B0015 with XbaI, after 2 hours at 37C I storaged the reactions at -20C.<br>
--Also I digested the T7* RNAP with XbaI and SpeI, I cleaned the BBa_B0015 from all the buffers & enzymes by column cleaning kit, and the concentration I got was 56ng/ul (9ul in total).
 ==Asaf==
 I cleaned the PCR products of the polymerase genes from the gel.
@@ Line 29: / Line 27: @@
 ==Shahar==
+-Clean the N$ & T3  RNAP after digestion by cleaning kit.
+<br> -Those inserts (N4 & T3 RNAP) are ready for cloning into pPROLAR plasmid.
 ==Rachel==
+-Clean the PCR product of K1F RNAP (the one with pPROLAR primers)from the gel. The concentration was: 17 ng/ul. <br>-Digest of K1F RNAP with HindIII and NHeI for creating sticky ends.<br>-Clean the K1F RNAP after digestio by cleaning kit. The final concentration: 25 ng/ul<br>-This insert (K1F RNAP) is ready for cloning into pPROLAR plasmid.
-}}