User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/08/18

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August 18th, 2010
1. Make colony PCR to verify 1) L1: p30-MinBP + GFP E0240 (6 colonies); 2) L2: p30-MinBP + GFP BBa_K145015 (74 min) (10 colonies); 3) L4: Backbone pSB3K3 + Blue Promoter BBa_K238013 + GFP BBa_K145015 (74 min) (3 colonies).
 * Tubes are marked as follows:
 * -2.1-2.10 -> L2: p30-MinBP + GFP BBa_K145015 (74 min) (10 colonies)
 * -4.1-4.3 -> L4: Backbone pSB3K3 + Blue Promoter BBa_K238013 + GFP BBa_K145015 (74 min) (3 colonies)


 * Primers used were Preffix FWD and Suffix REV, reactives needed for one reactions are as follows:


 * PCR with Taq DNA polymerase
 * Reactive (ul x sample)
 * Taq Polymerase -> 1
 * Taq Reaction Buffer 10X -> 5
 * MgCl 50mM (can be used up to 3ul) -> 2.5
 * dNTP’s 0.4ug/ul -> 2.5
 * Primer Forward (can be used up to 3ul) -> 2.5
 * Primer Reverse (can be used up to 3ul) -> 2.5
 * HPLC -> 24
 * DNA -> 10
 * Total volume -> 50


 * Thermocycler program:
 * 1. 95ºC 5 min
 * 2. 30 cycles
 * 95ºC 45 seg
 * 55ºC 45 seg
 * 72ºC 1.5 min
 * 3. 72ºC 10 min
 * 4. Hold 4ºC

2. Run gel to verify PCR made on August 13th and 18th.
 * FAILED!!!


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