IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/07/01
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The results of the PCR made yesterday are shown in the following electrophoresis gel(120V x 35min).
1. Ladder 5Kb. 2. PCR 1. 3. PCR 2. 4. PCR 3.
As expected the two first reactions corresponds to the fragment of intersts and the third which is the control showing nothing.
I mixed the two reactions into one and then I cleaned it with the High Purification PCR Products kit from Roche. The final volume was 50 microlitters and to see if nothing wrong happened I ran 3 microL in a Daniel's gel:
1. Ladder 1 Kb. 2. Daniel's stuff 1. 3. Daniel's stuff 2. 4. Daniel's stuff 3. 5. Clean PCR.
Digestion of the PCR
In order to repeat the procedure of the ligation by sticky ends of the PCR with pBBRMCS-5, I did again double digestion by apaI and sacI:
The ligation's proportions(microL):