From 2012.igem.org
Team:Technion/2_September_2012
Ilya
Inbal
-I ran the rest of T7* RNAP with pSB1A2 primers on agarose gel, but the band was too faded to purify it (still- I'm going to purify it, just in case...) so I repeat the PCR twice for it but now I changed the Tm, one reaction was Tm=54C, the other was Tm=56C (After consultation with Shachar and roee, we decided to change the Tm's to gradient tempratures). After running a 5ul sample from the PCR products, I got the desired bands (2700bp), with no other unwanted bands, So I purified the products by PCR cleaning kit. The concentrations are: for 54C:207ng/ul, for 56C: 298ng/ul.
- 3 starters of k133 (the inserts are pTetO+m-cherry) that I received from Roee's lab.
Asaf
I ran the PCR products of the polymerases on an extracting gel. I got the expected results (although N4 is rather week).
Hila
I wanted to restrict the fragments with PacI and SpeI, but…
NO PacI!!! :(
Lior
Noa
Colony PCR for: ALP + k1F, Sp6.
Miniprep and Nanodrop for all: xyIE + all, and ALP + N4, T3.
Evgeni
- For some reason I got no growth in starters, though there was growth on streak plate. Will try to make starters again, this time from the colonies on streak plate.
Shahar
PCR for N4 at 62 & 60 degrees, and for T3 worked with pPROLAR primers , but not with pTETO primers.
Digest of the RNAP with HindIII and NHeI for creating sticky ends
Rachel
I ran the PCR products of K1F RNAP (which designated for cloning into pPROLAR)on an extracting gel.
The result was as i expected for (2700 bp).
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