DNA extract
DNA extract kdorfman Mon, 03/12/2012 - 15:12Isolating, digest out the P-element, Ligate P-element line DNA
Use Qiagen DNeasy Blood & Tissue Kit 69504 $141.12 for 50 reactions
LiCl/KAc
LiCl/KAc kdorfman Mon, 03/12/2012 - 18:01LiCl/KAc
1 part 5M KAc : 2.5 parts 6M LiCl
Stock solution | 14 | 17.5 | 21 | mL final volume |
---|---|---|---|---|
5M KAc | 4 | 5 | 6 | mL |
6M LiCl | 10 | 12.5 | 15 | mL |
Ligase
Ligase kdorfman Mon, 03/12/2012 - 18:03T4 DNA Ligase
NEB M0202S
Get from New England Biolabs Freezer Program in Fernald
Store in freezer
Comes with 10X ligase buffer (includes ATP
NaOAc
NaOAc kdorfman Mon, 03/12/2012 - 18:063M NaOAc
PBS for DNA
PBS for DNA kdorfman Mon, 03/12/2012 - 18:04PBS
~200 mL/prep
4 preps per group
Aliquot 1 mL per group
Soution A
Soution A kdorfman Mon, 03/12/2012 - 18:00Solution A
100 mM Tris-HCl pH 7.5
100mM EDTA
100 mM NaCl
0.5% SDS
Stock solution | 10 | 25 | 50 | 100 | mL final volume |
---|---|---|---|---|---|
1 M Tris | 1 | 2.5 | 5 | 10 | mL |
0.5 M EDTA | 2 | 5 | 10 | 20 | mL |
5 M NaCl | 0.02 | 0.05 | 0.1 | 0.2 | mL |
20% SDS | 0.25 | 0.625 | 1.25 | 2.5 | mL |
Make 10 mL
pH to 7.5