IGEM:Cambridge/2008/Notebook/Magnetic Bacteria/2008/08/01

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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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MagA and I0500 Transformation Plates

 * MagA transformants grew on Ampicillin plates and Kanamycin plates but not Tetracyclin and Chloramphenicol
 * I0500 transformants grew on Kanamycin plates
 * pUC control also grew on Kanamycin plates and that's not good!
 * Suspected problems with preparation of our own Kanamycin plates and so re-do MagA, I0500 and pUC TOP10 transformants on Kanamycin plates prepared by Ellis
 * Plates incubated at 37°C overnight

Single Colony PCR of MagA from Ampicillin Plate

 * Without using master mix but normal protocol
 * Using updated PCR programme MAGA2
 * Touchdown: 6 cycles; Annealing max:56°C min:46°C
 * Normal: 28 cycles; annealing at 76°C
 * Reagents added in this order:
 * 12.4μL SDW
 * 4μL 5xHF Phusion Buffer
 * 0.4μL dNTPs (10mM)
 * 1μL primer MagAF (10mM)
 * 1μL primer MagAR (10mM)
 * 1μL DNA
 * 0.2μL Phusion DNA polymerase


 * Run on 0.8% agarose gel with SyBr Green
 * Colony 4 has a faint band at 1.3kb for MagA
 * Quickly run colonies 1 and 5 on E-gel and re-confirm the absence of MagA in these 2 colonies but not the fault of agarose gel




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