Noi/NOTES/2014-10-29: Difference between revisions

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= Generation of sequencing library of ''a customized SeqCap EZ Epi Probe Pool'' =
= 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

Back to calendar


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:
  1. If frozen, thaw the tube of SeqCap Epi probe pool on ice.
  2. Vortex the SeqCap Epi probe pool for 3 seconds.
  3. 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.
  4. 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.
  5. 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

  1. 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