Brandon:Protocols/Lysis buffers: Difference between revisions
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>Bsos m (→lysis buffer 1) |
>Bsos m (→lysis buffer 1) |
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Line 18: | Line 18: | ||
1000 uL of 1M NaCl | 1000 uL of 1M NaCl | ||
333 uL of 1M MgCl2 | 333 uL of 1M MgCl2 | ||
10 uL of NP40 (1% in soln, NP-40 substitute is fine) | |||
8.067 mL Nuclease free water | 8.067 mL Nuclease free water |
Revision as of 21:59, 1 May 2013
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% NP40
- make 1M solution of NaCl and MgCl2
1M NaCl2, 2.922 grams in 50 mL nuclease free water 1M MgCl2, 4.761 grams in 50 mL nuclease free water (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 10 uL of NP40 (1% in soln, NP-40 substitute is fine) 8.067 mL Nuclease free water