Brandon:Protocols/Lysis buffers: Difference between revisions

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*make 1M solution of NaCl and MgCl2
*make 1M solution of NaCl and MgCl2
  1M NaCl2, 2.922 grams in 50 mL nuclease free water
  1M NaCl2, 1.461 grams in nuclease free/DI water and bring to 25 mL
  1M MgCl2, 4.761 grams in 50 mL nuclease free water (add mgcl2 to water, very exothermic!!)
  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
*To make 10 mL of lysis buffer

Revision as of 20:01, 14 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% NP40
  • make 1M solution of NaCl and MgCl2
1M NaCl2, 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
10   uL of NP40 (1% in soln, NP-40 substitute is fine)
8.067 mL Nuclease free water