User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/06/23

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June 23rd, 2010
1. Run gel to verify PCR results from June 22nd.



Lanes: 1) Ladder; 2) Plat Taq p30-MinBP; 3) Plat Taq ~3 kb’s; 4) rTth p30-MinBP; 5) rTth Ctrl ~3 kb’s; 6) rTth p30-MinBP; 7) rTth Ctrl ~3 kb’s; 8) rTth Ctrl -; 9) Plat Taq p30-MinBP; 10) Plat Taq Ctrl ~3 kb’s; 11) Plat Taq Ctrl -; 12,13) Mariana’s samples.

2. Restrictions to GFP reporter (E0240) with XbaI and PstI and backbone plasmid (pSB3K3) with EcoRI and PstI.
 * Double restriction methods XbaI and PstI (5ul DNA, total volume 20 ul):
 * DNA -> 5ul
 * Buffer 4 -> 2ul (10% of total volume)
 * BSA (required by SpeI) -> 1ul
 * XbaI -> 1ul
 * SpeI -> 1ul
 * HPLC -> 10ul (to complete total volume of 20ul)
 * Incubate at 37ºC overnight


 * Double restriction methods EcoRI and PstI:
 * DNA -> 5ul
 * Buffer 4 -> 2ul (10% of total volume)
 * BSA (required by SpeI) -> 1ul
 * EcoRI -> 1ul
 * SpeI -> 1ul
 * HPLC -> 10ul (to complete total volume of 20ul)
 * Incubate at 37ºC overnight


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