Noi/NOTES/2012-5-16
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Probe production for the HiResChrPaint project
- Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Noi/NOTES/2012-5-14
- Since Dr. Zhang suggested that about 20% of the oligos having AP2 part should be fine. I will continue to digest the rest of amplified amplicons with DpnII and column purify
Probe IDs Conc. (ng/ul) Yields in 120ul (ug) Left over after trial (ug) #1-V4 83.70 10.04 5.00 #2-V4 125.10 15.01 12.00 #3-V4 110.10 13.21 13.20 #4-V4 133.80 16.06 8.00 #1-V6 122.50 14.70 14.70 #2-V6 106.20 12.74 12.74 #3-V6 132.20 15.86 15.86 #4-V6 101.70 12.20 12.20 DpnII digestion reaction set up
Probe IDs | Conc. (ng/ul) | Volume (ul) | H2O (ul) | 10X DpnII Buffer | DpnII | Total volume (ul) | # of tube |
#1-V4 | 83.70 | 59.74 | 22.26 | 10 | 8 | 100.00 | 1 |
#2-V4 | 125.10 | 47.96 | 34.04 | 10 | 8 | 100.00 | 2 |
#3-V4 | 110.10 | 59.95 | 22.05 | 10 | 8 | 100.00 | 2 |
#4-V4 | 133.80 | 59.79 | 22.21 | 10 | 8 | 100.00 | 1 |
#1-V6 | 122.50 | 60.00 | 22.00 | 10 | 8 | 100.00 | 2 |
#2-V6 | 106.20 | 59.98 | 22.02 | 10 | 8 | 100.00 | 2 |
#3-V6 | 132.20 | 59.98 | 22.02 | 10 | 8 | 100.00 | 2 |
#4-V6 | 101.70 | 59.98 | 22.02 | 10 | 8 | 100.00 | 2 |
37C 30min -> 65C 20min -> Qiaquick column purification (also combine the samples used for DpnII digestion trial. Measure concentration by Nanodrop and calculate yield