Noi/NOTES/2014-8-13
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Expansion PCR of new probe set from CustomArrays (90K_ologos_04Aug2014)[edit]
- Link to calendar
- 2014-08-13: Received MOMOD V4 Oligo (Dr. Zhang recently called V5) pool from CustomAraay
Oligo info.[edit]
- Name: 90K_ologos_04Aug2014
- Conc. 20.53ng/ul,
- Volume 80ul in TE buffer. Total amount 1.64ug
- Info from Dr. Zhang's note
Description # oligos Length Amp. Primers Adaptor Removal Set A: BSPP LMS cluster probes 73,574 148nt V6 USER/DpnII Set B: Selector LMS cluster 12,994 148nt V4 USER/DpnII Set C: Selector chr21 unique 2,539 148nt eMIP_CA_F/R Nt.AlwI/Nb.BrsDI Total: 89,107
- I will be working on 73K probe set
- Length 148 mer (MW = 148 * 330Da/bp = 48,840 Da or g/mole
- Conc. 20.53ng/ul = 420.35nM (20.53ng/ul/ 48,840g/mole)
Expansion PCR[edit]
95C 30sec -> (95C 30sec -> 54C 45sec-> 72C 45sec) X 16-> 72C 3min -> 15C hold
- -There is something wrong with the oligo pool since I can not amplify all the three subsets of the probes, but it worked for positive control of the three subsets.
- -We will get CustomArray to re-synthesize the probes