User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/08/19

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August 19th, 2010
1. Make PCR to amplify BluePromoter to re-do ligation L4: Backbone pSB3K3 + Blue Promoter BBa_K238013 + GFP BBa_K145015 (74 min). As control I used primers to amplify OGR gene.


 * Reactive (ul x sample)
 * Taq Polymerase -> 1
 * Taq Reaction Buffer -> 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 -> 32
 * DNA -> 2
 * Total volume -> 50


 * Thermocycler program:
 * 1. 95ºC 5 min
 * 2. 35 cycles
 * 95ºC 45 seg
 * 55ºC 45 seg
 * 72ºC 1 min
 * 3. 72ºC 10 min
 * 4. Hold 4ºC


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