User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/09/26

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September 26th, 2010
1. Run gel to verify PCR made on September 25th.



Lanes: 1) Green ladder; 2) pSB3K3-J23101 + Δ RBS-GFP E0040 (colony 1, tube 1); 3) pSB3K3-J23101 + Δ RBS-GFP E0040 (colony 2, tube 2); 4) pSB3K3-J23101 + Δ RBS-GFP E0040 (colony 3, tube 3); 5) pSB3K3-Min. Blue Promoter + Δ RBS-GFP E0040 (colony 1, tube 4).

2. Make the following restrictions:
 * SpeI-PstI double restriction to BBa_J61002 plasmid containing promoter J23101.
 * SpeI restriction to pSB3K3 + J23101.


 * SpeI-PstI double restriction methods:
 * DNA -> 5 ul
 * Buffer 2 -> 2 ul (10% of total volume)
 * BSA -> 1 ul
 * SpeI -> 1 ul
 * PstI -> 1 ul
 * HPLC -> 10 ul (to complete total volume of 20ul)
 * Incubate at 37º C for 4 hrs.


 * SpeI Restriction methods:
 * DNA -> 5ul
 * Buffer 4 -> 2ul (10% of total volume)
 * BSA -> 1ul
 * SpeI -> 1ul
 * HPLC -> 11ul (to complete total volume of 20ul)
 * Incubate at 37ºC 4 hrs.


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