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= Expansion PCR of new probe set from CustomArrays (90K_ologos_04Aug2014) = * [[noi:DMR220k_LabNotes| '''Link to calendar''']] * 2014-08-13: Received MOMOD V4 Oligo (Dr. Zhang recently called V5) pool from CustomAraay === Oligo info. === * 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 == 95C 30sec -> (95C 30sec -> 54C 45sec-> 72C 45sec) X 16-> 72C 3min -> 15C hold<br> :'''-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'''
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