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Team:Technion/23 August 2012
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-Planing the primers for the Gibson assembly. | -Planing the primers for the Gibson assembly. | ||
==Asaf== | ==Asaf== | ||
| + | I ran on gel agarose the PCR products of yesterday (the polymerase genes) for testing. <br> | ||
| + | I only got the expected band for T7 wild type. I suspect that the problem is with the miniprep process. | ||
==Hila== | ==Hila== | ||
| Line 34: | Line 36: | ||
==Evgeni== | ==Evgeni== | ||
| - | - Checking the plates after yesterday's transformation. Got many colonies on control plates meaning that many plasmids were cut in one site only... | + | - Checking the plates after yesterday's transformation. Got many colonies on control plates meaning that many plasmids were cut in one site only...<br> |
| + | -Putting a plate with transformed bacteria in 4C for starters preparation latter on. | ||
==Shahar== | ==Shahar== | ||
==Rachel== | ==Rachel== | ||
| - | + | Last chance for PCR to RNAP T7* - again no result at all. | |
| + | Now we have to plan and order new primers. | ||
}} | }} | ||
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Ilya
Inbal
- I made today 2 new starters of the BioBrick BBa_I13522.
-Planing new primers for the T7 RNAP from chris.
- Sending to sequence the pSB1C3+SP6 plasmid (Conc: 78 ng/ul).
-Planing the primers for the Gibson assembly.
Asaf
I ran on gel agarose the PCR products of yesterday (the polymerase genes) for testing.
I only got the expected band for T7 wild type. I suspect that the problem is with the miniprep process.
Hila
The bacteria grew on the negative control (no insert) plates… :(
Colony PCR for:
- Six positive colonies (pSB1C3+MCS)
- Two negative control colonies (pSB1C3 without insert?!)
- Positive control (pSB1C3 + Fus_1)
- Negative control (no colony inserted)
1% gel for the PCR products.
I got five positive colonies (number 1-4, 6) with the wanted 344 bp band.
Colony number 5 showed a 1106 bp band (consider negative for the ligation) like the negative control colonies and the positive control for the experiment.
We think that the CIP enzyme was contaminated so the reaction was not as good as it should…
No bend appeared in the negative control strip.
Lior
Colony PCR for xyIE: we got 1/5.
PCR purification for pT3, pN4, pSp6, we got high concentrations- yeah :)
Cutting of plasmis and inserts with XhoI and XmaI.
Noa
Evgeni
- Checking the plates after yesterday's transformation. Got many colonies on control plates meaning that many plasmids were cut in one site only...
-Putting a plate with transformed bacteria in 4C for starters preparation latter on.
Shahar
Rachel
Last chance for PCR to RNAP T7* - again no result at all. Now we have to plan and order new primers.
