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EPF-Lausanne/13 August 2008
From 2008.igem.org
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Molecular biology
Yesterday's cell cultures and mini-prep worked out fine.
F1610/E1010
We do the gel of F1610 and E1010 from the old mini-preps that we digested with SpeI and EcoRI all night. E1010 is great but F1610 doesn't work.
We don't know where the mistake comes from.
So we want to be sure that the part are correct and enzymes too. So we digest F1610 and E1010 from new mini-prep of today and we split experiences of digest in two EcoRI/SpeI and PstI/XbaI.
And we do different PCR by this way we don't use restriction enzyme. We do PCR with biobrick primer for E1010, F1610, F1610/E1010(assembled) and we do a positive control with DNA and Primers form Bart.
In finally we run a gel with :
F1610 digest by EcoRI and SpeI
E1010 digest by EcoRI and SpeI
F1610 digest by PstI and XbaI
E1010 digest by PstI and XbaI
F1610 from PCR
E1010 from PCR
Resultat
Restriction enzymes work. ???????????????????????
E1010/F1610 from PCR (2 times from different colonies)
Positiv control from PCR
E1010 (nothing, straight from mini-prep)
F1610 (nothing, straight from mini-prep)
I1466/I14033
The gel purification doesn't work. We don't see the insert, I1433. We focus on E1010 and F1610.
R../B0034
We focus on E1010 and F1610.
