Rui:Haplotying: Difference between revisions

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==Background==
==Background==


==Protocol==
Original: [[File:Design.ppt]]
Original: [[File:Design.ppt]]


Modified: [[File:Tn-based-haplotyping V1.1.ppt]]
Modified: [[File:Tn-based-haplotyping V1.1.ppt]]


*Cell lysis buffer (1*)
==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>