Brandon:Protocols/Lysis buffers: Difference between revisions
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>Bsos |
>Bsos m (→lysis buffers) |
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Line 8: | Line 8: | ||
100mM NaCl | 100mM NaCl | ||
30mM MgCl2 | 30mM MgCl2 | ||
1% NP40 | 1% IGEPAL® CA-630 (replaces NP40) | ||
*make 1M solution of NaCl and MgCl2 | *make 1M solution of NaCl and MgCl2 | ||
Line 18: | Line 18: | ||
1000 uL of 1M NaCl | 1000 uL of 1M NaCl | ||
333 uL of 1M MgCl2 | 333 uL of 1M MgCl2 | ||
100 uL of | 100 uL of IGEPAL® CA-630 (1% in soln, replaces NP40) | ||
8.067 mL Nuclease free water | 8.067 mL Nuclease free water |
Revision as of 01:01, 19 March 2014
lysis buffers
lysis buffer 1
- 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