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

From OpenWetWare

Jump to: navigation, search
iGEM UNAM-Genomics-Mexico Main project page
Previous entry      Next entry

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



Personal tools