Buffer composition for Qiagen MiniPrep, midiPrep, MaxPrep, PCR clean up and gel extraction kits

Buffer P1 – Resuspension Buffer
50mM Tris-Cl, pH 8.0, 10mM EDTA, 100ug/mL RNase A
Storage condition – 4oC after adding RNase A
Prep – Dissolve 6.06g Tris base, 3.72g EDTA-2H20 in 800mL dH20. Adjust the pH to 8.0 with HCl. Adjust the volume to 1 liter with dH2O. Add 100mg RNase A (Sigma R4642, or Qiagen #19051 and 19101) per liter of P1.

Buffer P2 – Lysis Buffer
200mM NaOH, 1% SDS
Store at RT
Dissolve 8.09g of NaOH pellets in 950mL dH2O, 50mL 20% SDS solution. The final volume should be 1 liter.

Buffer P3 – Neutralization Buffer for Qiatips, Midiprep, Maxiprep, and Gigaprep kits.
3.0M potassium acetate, pH 5.5
Store at 4oC or RT
Dissolve 294.5g potassium acetate in 500mL dH2O. Adjust the pH to 5.5 with glacial acetic acid (about 110mL). Adjust the volume to 1 liter with dH2O.

Buffer N3 – Neutralization Buffer for spin columns.
4.2 M Gu-HCl, 0.9 M potassium acetate, pH 4.8

Buffer PE – Wash Buffer
10 mM Tris-HCl pH 7.5, 80% ethanol Store at RT

Buffer QBT - Equilibrium Buffer
750mM NaCl, 50mM MOPS, pH7.0, 15% isopropanol, 0.15% Triton X-100
Store at RT
Dissolve 43.83g NaCl, 10.46g MOPS (free acid) in 800mL dH2O. Adjust the pH to 7.0. Add 150mL pure isopropanol and 15mL 10% Triton X-100 solution. Adjust the volume to 1 liter with dH2O.

Buffer QC – Wash Buffer
1.0M NaCl, 50mM MOPS, pH 7.0, 15% isopropanol
Store at RT
Dissolve 58.44g NaCl and 10.46g MOPS (free aicd) in 800mL dH2O. Adjust the pH to 7.0. Add 150mL pure isopropanol. Adjust the volume to 1 liter with dH2O.

Buffer QF – Elution Buffer
1.25M NaCl, 50mM Tris-Cl, pH 8.5, 15% isopropanol
Store at RT
Dissolve 73.05g NaCl and 6.06g Tris base in 800mL dH2O and adjust the pH to 8.5 with HCl. Add 150mL pure isopropanol. Adjust the volume to 1 liter with dH2O.

Buffer QN
1.6M NaCl, 50 mM MOPS pH 7.0, 15% isopropanol

Buffer FWB2
1M potassium acetate, pH 5.0

TE Buffer
10mM Tris-Cl, pH 8.0, 1mM EDTA
Store at RT

STE Buffer
100mM NaCl, 10mM Tris-Cl, pH 8.0, 1M EDTA
Store at RT
Dissolve 5.84g NaCl, 1.21g Tris base and 0.37g EDTA-2H2O in 800mL dH2O and adjust the pH to 8.0 with HCl. Add up the volume to 1 liter with dH2O.

EB Buffer – Elution Buffer
10mM Tris-Cl, pH 8.5
Store at RT

QG Buffer – Solubilization Buffer for Gel Extration Kit
Composition Unknown (Proprietary)
Store at RT

Home-made QG Buffer

Recipe 1
90.8 g NaI
1.5 g Na2SO3
100 ml of water.
Filter through 0.45 um filter,
then add another 0.5 g Na2SO3.
Store in the coldroom in a dark bottle.

Recipe 2
6M Guanidine thiocyanate
50mM Tris-HCl pH 7.5
20mM EDTA pH 8.0

Recipe 3
60% Guanidine thiocyanate (5.36M)
140mM MES (2-[N-Morpholino] ethanesulfonic acid)
0.006% Phenol Red

PB Buffer - Binding Buffer
5 M Gu-HCl, 30% isopropanol
Store at RT

July 14, 2010 at 1:14 pm

1 Comment »

  1. DK said,

    August 6, 2011 @ 6:28 pm

    Buffer PB composition is wrong:
    1) 30% isopropanol, not ethanol
    2) no tris or any other buffer (and, seriously, why would anyone use tris to buffer at pH 6.6???)

    http://methodsandreagents.pbworks.com/w/page/20806948/Qiagen%27s%20Miniprep%20buffers

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