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Team:Technion/30 August 2012
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- 2 starters for the biobrick BBa_K346000 and pACSP6R (there were few colonies on the plates from yesterday), the plates are stored at 4C. <br> | - 2 starters for the biobrick BBa_K346000 and pACSP6R (there were few colonies on the plates from yesterday), the plates are stored at 4C. <br> | ||
-PCR purification for T7* RNAP (the one with the PROLAR primers), the conc.:114ng/ul.<br> | -PCR purification for T7* RNAP (the one with the PROLAR primers), the conc.:114ng/ul.<br> | ||
| - | -PCR for T7* RNAP with the pSB1A2 primers, Tm of 55C with elongation time of 3 minutes. The results were AWSOME- I got 2700 bp band! (the expected size!), but there were a smear above 2700bp and primer dimers at the bottom of the gel- So I need to seperate and purify the 2700 band (I will do it | + | -PCR for T7* RNAP with the pSB1A2 primers, Tm of 55C with elongation time of 3 minutes. The results were AWSOME- I got 2700 bp band! (the expected size!), but there were a smear above 2700bp and primer dimers at the bottom of the gel- So I need to seperate and purify the 2700 band (I will do it on Sunday).<br> |
-mini prep of the starters from the morning- the concentrations are: for BBa_K346000: 218ng/ul, for pACSP6R: 141ng/ul. | -mini prep of the starters from the morning- the concentrations are: for BBa_K346000: 218ng/ul, for pACSP6R: 141ng/ul. | ||
==Asaf== | ==Asaf== | ||
Revision as of 19:12, 5 September 2012
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Ilya
Inbal
- 2 starters for the biobrick BBa_K346000 and pACSP6R (there were few colonies on the plates from yesterday), the plates are stored at 4C.
-PCR purification for T7* RNAP (the one with the PROLAR primers), the conc.:114ng/ul.
-PCR for T7* RNAP with the pSB1A2 primers, Tm of 55C with elongation time of 3 minutes. The results were AWSOME- I got 2700 bp band! (the expected size!), but there were a smear above 2700bp and primer dimers at the bottom of the gel- So I need to seperate and purify the 2700 band (I will do it on Sunday).
-mini prep of the starters from the morning- the concentrations are: for BBa_K346000: 218ng/ul, for pACSP6R: 141ng/ul.
Asaf
I tried again the PCR of the 3 polymerase genes that failed the previues PCR (T7*,N4,K1F), now with Tm of 66C.
I tried this Tm because I got results with this Tm before (29.8 polymerase gel 2). This time I got it right,
I got the 2.7 kb bands I was expecting (althouge N4 is rather week).
 |
Hila
Lior
Noa
olony PCR for XyIE bacteria described on 28.
Positive results for all strains!! yeah :)
Evgeni
- Transformation of TOP10 RB cells with pPROLar+Bba_015 ligated part
- Plating of transformed cells on Kanamycin plates - 100 ul and rest
Shahar & Rachel
We tried to do the failed PCR's from yesterday, but with lower temprature. For N4 (with A2 & plac/ara primers)we tried with temprature gradient.
