Jie:LabNotes/CpgSeq/2009-8-26: Difference between revisions
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>Sam Chiang No edit summary |
>Jie deng No edit summary |
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96C 3min-> 45x (95C 30sec, MATN4 and DNMT3B at 58C, rs10846023 on 62C 1min, 72C 1min)-> 72C 5min. | 96C 3min-> 45x (95C 30sec, MATN4 and DNMT3B at 58C, rs10846023 on 62C 1min, 72C 1min)-> 72C 5min. | ||
==TA cloning of the above 8 samples== | |||
Step 1. Cloning into pCR2.1. | |||
Set up the 10ul ligation reaction as follows: | |||
x 6 | |||
DNA template 0.5ul 3 | |||
10x Ligation Buffer 0.5ul 3 | |||
pCR2.1 vector(25ng/ul) 1ul 6 | |||
water 2.5ul 15 | |||
T4 DNA ligase(4 Weiss Units)0.5ul 3 | |||
Incubate the ligation reaction at 14C overnight. |
Revision as of 18:28, 27 August 2009
ASM validation of cell type and individual dependent ASM
primers for cell and individual dependent ASM validation
I did the PCR using 2x iQ supermix on: rs10846023_FLJ22662 on BJ(7/26/09) and BJiPS11(; MATN4 on PGP1F(10/10/08) and PGP1iPS; DNMT3B on PGP3L and PGP9L; x 7 2x iQ supermix 50 350 Primer F (50uM) 0.5 0.5 each Primer R (50uM) 0.5 0.5 each sample bis-gDNA (~202ng/ul) 0.1 0.1 each dH2O 50 350 total 40ul
96C 3min-> 45x (95C 30sec, MATN4 and DNMT3B at 58C, rs10846023 on 62C 1min, 72C 1min)-> 72C 5min.
TA cloning of the above 8 samples
Step 1. Cloning into pCR2.1.
Set up the 10ul ligation reaction as follows: x 6 DNA template 0.5ul 3 10x Ligation Buffer 0.5ul 3 pCR2.1 vector(25ng/ul) 1ul 6 water 2.5ul 15 T4 DNA ligase(4 Weiss Units)0.5ul 3 Incubate the ligation reaction at 14C overnight.