Brandon:Protocols/Lysis buffers: Difference between revisions

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>Bsos
>Bsos
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, NP-40 substitute is fine)
  100  uL of NP40 (1% in soln, IGEPAL® CA-630 substitute used)
  8.067 mL Nuclease free water
  8.067 mL Nuclease free water

Revision as of 22:52, 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 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, IGEPAL® CA-630 substitute used)
8.067 mL Nuclease free water