Brandon:Protocols/Lysis buffers

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lysis buffers[edit]

lysis buffer 1[edit]

  • 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
' ' 10X 100X 50X 40X '
Tris-HCl pH 7.5 2.0M 500 5000 2500 2000 uL
NaCl 5M 200 2000 1000 800 uL
MgCl 1M 333 3330 1665 1332 uL
IGEPAL CA-630 solid 100 1000 500 400 uL
0 0 0 uL
11330 5665 4532 uL
H20 -1330 4335 5468 uL