Matt:LabNotes/2013-3-9
Jump to navigation
Jump to search
Second Strand cDNA synthesis[edit]
- Follow NEB E6111S protocol: [1]
Started with 20 ul cDNA from Friday 3/8/13
- Put in ice
- Added 48 ul H20
- Added 8ul 10x Second strand synthesis reaction buffer
- Added 4ul Second strand synthesis enzyme mix
- Mixed by pipetting
- Incubated for 2.5 hours at 16C
- Purified using Qiagen minelute column (eluted 23ul into 1.5ml tube labelled HBRR cDNA)
- Measured DNA conc with Nandrop: 55.5 ng/ul
- 21.5ul (1.19ug) sample left in tube