From 2012.igem.org
Team:Technion/9_September_2012
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.
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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
Me and Noa did restriction for the I13600 part and than we made ligation with xyIE and ALP and transformation to Top10 cells.
Noa
Evgeni
- Restriction analysis of starters with NheI and also with BamHI+HindIII. Both analysis are negative - they show no insert.
Shahar
Rachel
-3 Ligations: pCP+fus2, pCP+fus2del, pCP without insert (control).
- Transformation after ligations.
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