Rui:Haplotying: Difference between revisions
Jump to navigation
Jump to search
>RuiLiu |
>RuiLiu No edit summary |
||
Line 1: | Line 1: | ||
==Background== | ==Background== | ||
Original: [[File:Design.ppt]] | Original: [[File:Design.ppt]] | ||
Modified: [[File:Tn-based-haplotyping V1.1.ppt]] | Modified: [[File:Tn-based-haplotyping V1.1.ppt]] | ||
==Protocol== | |||
Step 1: Cell lysis | |||
Collect 10 cells in 20ul lysis buffer. | |||
Cell lysis buffer (1*) | |||
200mM NaCl | 200mM NaCl | ||
20mM EDTA | 20mM EDTA | ||
Line 12: | Line 17: | ||
0.2% TritonX-100 | 0.2% TritonX-100 | ||
(0.1 AU/ml Qiagen Proteinase 7.5AU) | (0.1 AU/ml Qiagen Proteinase 7.5AU) | ||
Step 2: Proteinase and aliquot | |||
Add Proteinase right before aliquot, 1ul per tube, to 20 tubes. Store at -80C. | |||
Step 3: Tagmentation | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''gDNA''' | |||
| align="center" style="background:#f0f0f0;"|'''1ul''' | |||
|- | |||
| HMW buffer||1ul | |||
|- | |||
| transposomes||1ul | |||
|- | |||
| Nf water||2ul | |||
|- | |||
| Total ||5ul | |||
|} | |||
==Primers== | ==Primers== |
Revision as of 23:18, 16 March 2011
Background
Original: File:Design.ppt
Modified: File:Tn-based-haplotyping V1.1.ppt
Protocol
Step 1: Cell lysis
Collect 10 cells in 20ul lysis buffer.
Cell lysis buffer (1*) 200mM NaCl 20mM EDTA 10mM Tris-HCl (pH 8) 0.2% TritonX-100 (0.1 AU/ml Qiagen Proteinase 7.5AU)
Step 2: Proteinase and aliquot
Add Proteinase right before aliquot, 1ul per tube, to 20 tubes. Store at -80C.
Step 3: Tagmentation
gDNA | 1ul |
HMW buffer | 1ul |
transposomes | 1ul |
Nf water | 2ul |
Total | 5ul |
Primers
File:Primers for Tn-based-haplotyping.xls
2011
<calendar> name=Rui:LabNotes/Haplotyping format=%name/%year-%month-%day date=2011/10/1 view=oneyear </calendar>