Brandon:Protocols/Lysis buffers: Difference between revisions

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>Bsos
>Bsos
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 NP40 (1% in soln, IGEPAL® CA-630 used instead)
  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