Brandon:Protocols/Lysis buffers
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lysis buffers[edit]
lysis buffer 1[edit]
- Crawford et al. PNAS 2003
- lysis buffer composition:
10X lysis buffer 100mM Tris.Hcl pH 7.5 100mM NaCl 30mM MgCl2 1% IGEPAL® CA-630 (replaces NP40)
- make 1M solution of NaCl and MgCl2
1M NaCl, 1.461 grams in nuclease free/DI water and bring to 25 mL 1M MgCl2, 2.3805 grams in nuclease free/DI water and bring to 25 mL (add mgcl2 to water, very exothermic!!)
- To make 10 mL of lysis buffer
500 uL of 2M Tris.Hcl PH7.5 1000 uL of 1M NaCl 333 uL of 1M MgCl2 100 uL of IGEPAL® CA-630 (1% in soln, replaces NP40) 8.067 mL Nuclease free water
' | ' | 10X | 100X | 50X | 40X | ' |
Tris-HCl pH 7.5 | 2.0M | 500 | 5000 | 2500 | 2000 | uL |
NaCl | 5M | 200 | 2000 | 1000 | 800 | uL |
MgCl | 1M | 333 | 3330 | 1665 | 1332 | uL |
IGEPAL CA-630 | solid | 100 | 1000 | 500 | 400 | uL |
0 | 0 | 0 | uL | |||
11330 | 5665 | 4532 | uL | |||
H20 | -1330 | 4335 | 5468 | uL | ||