Matt:LabNotes/2015-1-12

From ZhangLabWiki
Revision as of 23:19, 12 January 2015 by >Mzcai (Created page with "==Make First Strand cDNA for in vitro Capture with CA12k_Nov2014 Probe Set== * Followed NEB E6300S protocol: [https://www.neb.com/protocols/1/01/01/first-strand-cdna-synthesis...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

Make First Strand cDNA for in vitro Capture with CA12k_Nov2014 Probe Set

  • Followed NEB E6300S protocol: [1]
  • 2 tubes for RNA sample and 1 tube with no RNA (replace with H2O) as negative control
Components Volume
RNA 1.5 ul (0.75 ug)
d(T)23VN (50 µM) 2 ul
H2O 4.5 ul
Total 8 ul
  • Denatured RNA for 5 min at 70C and then put on ice
  • Added the following to tubes
Components Volume
M-MuLV Reaction Mix 10 ul
M-MuLV Enzyme Mix 2 ul
Total 20 ul
  • Incubated for one hour at 42C
  • Inactivated enzyme for 5 min at 80C