Noi/NOTES/2014-10-29: Difference between revisions
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>Noi (Created page with "= Generation of sequencing library of ''a customized SeqCap EZ Epi Probe Pool'' = '''Back to calendar''' * 2014-10-29: I am expecting the package...") |
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= Generation of sequencing library of | = Generation of sequencing library of a customized SeqCap EZ Epi Probe Pool = | ||
[[noi:DMR220k_LabNotes#2014|'''Back to calendar''']] | [[noi:DMR220k_LabNotes#2014|'''Back to calendar''']] | ||
* 2014-10-29: I am expecting the package of a customized SeqCap EZ Epi probe pool that Dr. Zhang ordered for target enrichment of the blood UMRs from all WGBS libraries | * 2014-10-29: I am expecting the package of a customized SeqCap EZ Epi probe pool that Dr. Zhang ordered for target enrichment of the blood UMRs from all WGBS libraries |
Revision as of 09:34, 29 October 2014
Generation of sequencing library of a customized SeqCap EZ Epi Probe Pool
- 2014-10-29: I am expecting the package of a customized SeqCap EZ Epi probe pool that Dr. Zhang ordered for target enrichment of the blood UMRs from all WGBS libraries
Things to do upon receive the probe pool
1. Recommended by instruction in CHAPTER2:
- Upon receipt, undertake the following steps to ensure the highest performance of the SeqCap Epi probe pool to avoid multiple freeze/thaw cycles or potential accidental contamination:
- If frozen, thaw the tube of SeqCap Epi probe pool on ice.
- Vortex the SeqCap Epi probe pool for 3 seconds.
- Centrifuge the tube of SeqCap Epi probe pool at 10,000 x g for 30 seconds to ensure that the liquid is at the bottom of the tube before opening the tube.
- Aliquot the SeqCap Epi probe pool into single-use aliquots (4.5 µl/aliquot) in 0.2 ml PCR tubes (or 96-well plates if following the higher throughput protocol described in Appendix C) and store at -15 to -25°C until use. The presence of some residual volume after dispensing all single-use aliquots is normal.
- When ready to perform the experiment, thaw the required number of single-use SeqCap Epi probe pool aliquots on ice.
NOTE: The SeqCap Epi probe pool should not undergo multiple freeze/thaw cycles. To help ensure the highest performance of the SeqCap Epi probe pool, Roche NimbleGen recommends aliquoting the SeqCap Epi probe pool into single-use volumes to prevent damage from successive freeze/thaw cycles.
Label the 8-tube strip: UMRs EZ Epi probe, Date 141029 Location: -20C, in small zip lock bag within Noi Library prep kit box
2. For sequencing library preparation
- Take a small aliquot the left over of the probe pool after a single-use aliquots for 12 rxn for library prep using ClonTech DNA SMRT kit