Brandon:Protocols/Lysis buffers
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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 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 NP40 (1% in soln, NP-40 substitute is fine) 8.067 mL Nuclease free water