"
Team:Technion/30 August 2012
From 2012.igem.org
Liorlevyzzz (Talk | contribs) |
|||
| (16 intermediate revisions not shown) | |||
| Line 4: | Line 4: | ||
==Inbal== | ==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. <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 on Sunday).<br> | ||
| + | -mini prep of the starters from the morning- the concentrations are: for BBa_K346000: 218ng/ul, for pACSP6R: 141ng/ul. | ||
==Asaf== | ==Asaf== | ||
I tried again the PCR of the 3 polymerase genes that failed the previues PCR (T7*,N4,K1F), now with Tm of 66C.<br> | I tried again the PCR of the 3 polymerase genes that failed the previues PCR (T7*,N4,K1F), now with Tm of 66C.<br> | ||
I tried this Tm because I got results with this Tm before (29.8 polymerase gel 2). This time I got it right,<br> | I tried this Tm because I got results with this Tm before (29.8 polymerase gel 2). This time I got it right,<br> | ||
| - | I got the 2.7 kb bands I was expecting (althouge N4 is rather week). | + | I got the 2.7 kb bands I was expecting (althouge N4 is rather week).<br> |
| - | Image:30. | + | <gallery perrow="4" widths=150px heights=150px style="color: black; border: 5px ridge #A7A3BF; font: 13px david; background: azure"> |
| + | Image:30.8_polymerase_test_gel4-view.jpg | ||
| + | </gallery> | ||
==Hila== | ==Hila== | ||
| Line 15: | Line 20: | ||
==Noa== | ==Noa== | ||
| + | colony PCR for XyIE bacteria described on 28. <br> | ||
| + | Positive results for all strains!! yeah :) | ||
==Evgeni== | ==Evgeni== | ||
Latest revision as of 11:53, 17 September 2012
- Home
- Team
- Project
- Parts
- Notebook
- Safety
- Attributions & Achievements
- Human Practices
- Sponsors & Thanks
|
|
|
|
|
|
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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
colony 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.
