IGEM:UNAM LCG/2009/Notebook/Hydrobium etli/2011/09/20
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pBBR1MCS backbone PCR extraction
I made a PCR to extract the backbone of pBBR1MCS. The plasmid is standarized so I used the prefix fwd and suffix reverse primers. I need this backbones to insert the entire Anderson collection in R. etli. I used the plasmids extracted by Vladimir (see the previous entrance). I ran a gel (on 21/09/2011) to see if the PCRs were well, but unfourtunatly nothing appeared.
I ran PCRs from the 4 plasmid extractions made by Vladimir to extract the backbone. I used the Invitrogen's™ Platinum® Taq DNA polymerase, to define the reactives consentrations and PCR steps I checked out the manual online for the enzyme: PCR taq platinum manual.
I made 4 reactions, one for eachplasmid extraction made by Vladimir: pBBR1MCS 1, pBBR1MCS 2, pBBR1MCS DH5α and pBBR1MCS S17.