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)
  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