Matt:LabNotes/2016-4-11: Difference between revisions
Jump to navigation
Jump to search
>Mzcai (Created page with " #Wash with nf-H2O twice #Prepare Ampligase mix on ice #*Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme *Combine B11 and B12...") |
>Mzcai mNo edit summary |
||
Line 5: | Line 5: | ||
#*Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme | #*Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme | ||
*Combine B11 and B12: 100ul at ~200nM | *Combine B11 and B12: 100ul at ~200nM | ||
**B12 looked to have some very fine PA gel still in there (not filtered enough?) | |||
{| {{table}} | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''Component''' | | align="center" style="background:#f0f0f0;"|'''Component''' |
Revision as of 01:38, 12 April 2016
- Wash with nf-H2O twice
- Prepare Ampligase mix on ice
- Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme
- Combine B11 and B12: 100ul at ~200nM
- B12 looked to have some very fine PA gel still in there (not filtered enough?)
Component | Volume |
DEPC-H2O | 47 |
Ampligase Buffer | 10 |
Batch 11 & 12 Padlock Probes ~200nM]] | 33 |
Ampligase | 10 |
Total | 100 |
- Add mix to sample and incubate for 30min at 37C
- Move sample to 60C and oven slowly decreases to 55C and held for ~24hr