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EPF-Lausanne/14 August 2008
From 2008.igem.org
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Molecular biology
We do a mini-prep of E1010, "F1610" and P0140
E1010/F1610
Yesterday we found out that F1610 has a size of 300bp, instead of 800bp. So there are two hypotheses: 1) The mini-prep is bad or 2) the biobrick from IGEM is bad.
So we do another digestion series with EcoRI and SpeI. We do it with different mini-preps of F1610 and with the last mini-prep of E1010 that worked (as a positive control).
Some mini-preps of F1610 haven't enough liquid left, so we do a PCR with these (we use E1010 from the last mini-prep as a positive control).
At the same time we do a PCR of all plasmids that should have E1010/F1610 ligated to see if there is a fragment of 1500 bp.
Result of Gel
All mini-preps of F1610 have a fragment of 300 bp. So the biobrick from IGEM doesn't work. No ligated fragment E1010/F1610 is visible.
(see Gel picture on page 75 in lab notebook)
Test of specific primer of BBs
We do a PCR to see if the specific primers that Alexandre ordered work with our biobricks. We will do the gel tomorrow.
