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Team:Technion/2 September 2012
From 2012.igem.org
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==Inbal== | ==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 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.<br> | + | -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.<br> |
- 3 starters of k133 (the inserts are pTetO+m-cherry) that I received from Roee's lab. | - 3 starters of k133 (the inserts are pTetO+m-cherry) that I received from Roee's lab. | ||
Revision as of 19:22, 5 September 2012
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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==
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.
