AlanFung:LabNotes/CTCF/2011-6-30: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 |
>Alan6017518 |
||
Line 22: | Line 22: | ||
==Protocol== | ==Protocol== | ||
==Proteinase K/Protease Digestion== | ==Proteinase K/Protease Digestion== | ||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Content''' | |||
| align="center" style="background:#f0f0f0;"|'''A''' | |||
| align="center" style="background:#f0f0f0;"|'''B''' | |||
| align="center" style="background:#f0f0f0;"|'''C''' | |||
| align="center" style="background:#f0f0f0;"|'''D''' | |||
| align="center" style="background:#f0f0f0;"|'''E''' | |||
| align="center" style="background:#f0f0f0;"|'''F''' | |||
|- | |||
| M-Digestion Buffer (2X)||10||10||10||10||10||10 | |||
|- | |||
| Sample||1.2||1.2||1.2||1.2||0||0 | |||
|- | |||
| 1:10 Proteinase K||1||0||0||0||0||0 | |||
|- | |||
| Proteinase K||0||1||0||0||1||0 | |||
|- | |||
| Protease||0||0||0||1||0||0 | |||
|- | |||
| UV-RT PCR Grade Water||7.8||7.8||8.8||7.8||9||10 | |||
|- | |||
| Total||20||20||20||20||20||20 | |||
|} |
Revision as of 19:04, 30 June 2011
N9-MDA Bisulfite Converted Sequencing Library Construction
Objective
- Figure out source of contamination from 6/28 experiment
- Repeat 6/28 experiment with only gDNA (UV EB buffer, use RT-PCR grade water as NTC)
- Sample List
- A-600pg gDNA W 0.1ul proteinase K
- B-600pg gDNA W proteinase k
- C-600pg gDNA W/O proteinase k
- D-600pg gDNA W protease
- E-NTC W proteinase K
- F-NTC W/O proteinase K
Overview
- Proteinase K/Protease Digestion
- Bisulfite Conversion
- Heat Denature
- MDA
- Ethanol Precipitation
- Nextera Library Construction (Low input protocol)
- PAGE Analysis
Protocol
Proteinase K/Protease Digestion
Content | A | B | C | D | E | F |
M-Digestion Buffer (2X) | 10 | 10 | 10 | 10 | 10 | 10 |
Sample | 1.2 | 1.2 | 1.2 | 1.2 | 0 | 0 |
1:10 Proteinase K | 1 | 0 | 0 | 0 | 0 | 0 |
Proteinase K | 0 | 1 | 0 | 0 | 1 | 0 |
Protease | 0 | 0 | 0 | 1 | 0 | 0 |
UV-RT PCR Grade Water | 7.8 | 7.8 | 8.8 | 7.8 | 9 | 10 |
Total | 20 | 20 | 20 | 20 | 20 | 20 |