Daniel:Notebook/ComboLock/2016-6-24: Difference between revisions
Jump to navigation
Jump to search
>Djacobse (Created page with "=Stage 2 Test (Started yesterday) Back to Calendar ==Protocol== <ol start="2"> <li>Latch and Padlock H...") |
>Djacobse No edit summary |
||
Line 1: | Line 1: | ||
=Stage 2 Test (Started [[Daniel:Notebook/ComboLock/2016-6-23|yesterday]]) | =Stage 2 Test (Started [[Daniel:Notebook/ComboLock/2016-6-23|yesterday]])= | ||
[[Daniel:Notebook/ComboLock|Back to Calendar]] | [[Daniel:Notebook/ComboLock|Back to Calendar]] |
Revision as of 22:56, 22 June 2016
Stage 2 Test (Started yesterday)
Protocol
- Latch and Padlock Hybridization
- Heat inactivate enzyme by incubating for 2 minutes at 94C
- Exonuclease Digestion
- Prepare exonuclease I/III mix by mixing exonuclease I (20 units/μL) and exonuclease III (200 units/μL) in 1:1 ratio
- Add 2 μL of exonuclease I/III mix to reaction
- Mix the reaction by swirling pipette around the well 5 times
- Incubate reaction at 37 ºC for 2 hours
- Heat inactivate enzyme by incubating at 94C for 5 minutes
- qPCR
- Set up reaction according to table
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x20
- 72C 2 min
- 16C hold