Team:Technion/9 September 2012
From 2012.igem.org
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==Inbal== | ==Inbal== | ||
- | + | -PCR for pTetO+m-cherry: the PCR went well and I got a band at 800bp (expected), then I cleaned the PCR products (concentration: 17 ng/ul). <br> | |
+ | -6 starters for T7*+terminator+pSB1AK3.<br> | ||
+ | -Digestion of native T3 RNAP with XbaI and SpeI. running on gel and purifing the desired band (2700bp) from the plasmid backbone (2070bp), the concentration was 40.5ng/ul. | ||
+ | - Also, I checked if SP6 RNAP gene is within pSB1C3 by digesting it with PstI, after running the products on an agarose gel, I saw faded band at 3kb (not expected), and a small smear above it... I decided to let go of the SP6 for now.<br> | ||
+ | - | ||
==Asaf== | ==Asaf== | ||
I ran the PCR products of the fusion between the Riboswitch and the different polymerase genes (without K1F) on a gel.<br> | I ran the PCR products of the fusion between the Riboswitch and the different polymerase genes (without K1F) on a gel.<br> |
Revision as of 14:13, 15 September 2012
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Ilya
Inbal
-PCR for pTetO+m-cherry: the PCR went well and I got a band at 800bp (expected), then I cleaned the PCR products (concentration: 17 ng/ul).
-6 starters for T7*+terminator+pSB1AK3.
-Digestion of native T3 RNAP with XbaI and SpeI. running on gel and purifing the desired band (2700bp) from the plasmid backbone (2070bp), the concentration was 40.5ng/ul.
- Also, I checked if SP6 RNAP gene is within pSB1C3 by digesting it with PstI, after running the products on an agarose gel, I saw faded band at 3kb (not expected), and a small smear above it... I decided to let go of the SP6 for now.
-
Asaf
I ran the PCR products of the fusion between the Riboswitch and the different polymerase genes (without K1F) on a gel.
As you can see I got no results.
I thought that the problem may be the Tm, so I did a fusion PCR of the Riboswitch with the different polymerase genes
(without K1F) in a grad PCR with Tm of 55C and 56C.
Hila
Lior
Noa
Evgeni
- Restriction analysis of starters with NheI and also with BamHI+HindIII. Both analysis are negative - they show no insert.