From 2012.igem.org
Team:Technion/6_September_2012
Ilya
Inbal
- Transformation after ligation of T7*+ter (within pSB1AK3 plasmid) and control (plasmid after cip treatment)to the TOP10 component cells. I plated 100ul and rest on AMP antibiotics plates.
Asaf
Today I tried the fusion PCR of the RiboSwitch with the K1f polymerase gene with a lower gradient temperture (55,56,57C).
I did it for greater resulution.
I ran the PCR pruducts on agarose gel to test them. I got good results. I can see clearly the 3.2kb bands, the other
bands are for the K1F polymerase (2.7kb) and for the RiboSwitch (500bp).
Because I found the right Tm, I used Tm=57C for the fusion PCR of the Riboswitch with the rest of the polymerases genes.
Hila
Lior
We minipreped the I13600 part we had in the glycerol stock and we did PCR for the part.
Noa
Evgeni
- Transformation result: I had seen growth only on rest plates, few colonies on control plate. I've put plates in 4C
- Centrifugation and freezing of pPROLar starters
Shahar
miniprepd N4 Rnap from chris. preformed PCR with new templates at temprature 54-56 degrees (with pTETO primers).
This time it worked with 54 & 55 degrees.
Rachel
- miniprep of pCP plasmid.
- Restriction digest of the pCP backbone with XhoI and XbaI.
- Ran the pCP restriction products on a gel. I got the desired bands (2553bp), So I purified the products by PCR cleaning kit. The concentration was 15.4 ng/ul.
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