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Alberta NINT/18 August 2008
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[[Team:Alberta_NINT/Notebook | < Back to notebook]] | [[Team:Alberta_NINT/Notebook | < Back to notebook]] | ||
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4 C hold | 4 C hold | ||
| - | + | lab work (JD): | |
| + | Glycerol stock of K102042, K102052, K102062 and K102072 | ||
| + | Transformed XL1-B cells with K102041, K102051, K102061, K102071 | ||
| + | Co-transformed XL1-B cells with K102041+K102042, K102051+K102052, K102061+K102062, K102071+K102072 | ||
| + | Plated transformed cells and incubated overnight | ||
Latest revision as of 23:09, 11 October 2008
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lab work (SD):
Extracted pSB1A3.1 and pSB1A3.3 from the gel from 15/08/08 using QIAquick gel extraction protocol.
Incubated at 65 C for 8 minutes, vortexed twice for 10 sec. Eluted in 30ul of Buffer EB and let
stand for 2 min prior to centrifuging to increase the concentration.
Sequenced TA2.1, TA2.7, TA3.1, TA3.2, TA6.1 and TA7.2 again using an adapted sequencing protocol.
Combined DNA, Vf primer, Sequencing buffer, Pellet Paint and MQ H20, heated at 98 C for 5 min.
Added Big Dye mix and cooled on ice for 2 min.
Proceeded with sequencing:
[96 C for 10 sec
50 C for 20 sec
60 C for 4 min] X30
4 C hold
lab work (JD):
Glycerol stock of K102042, K102052, K102062 and K102072
Transformed XL1-B cells with K102041, K102051, K102061, K102071
Co-transformed XL1-B cells with K102041+K102042, K102051+K102052, K102061+K102062, K102071+K102072
Plated transformed cells and incubated overnight
