DNA purification buffers, mostly copied from OpenWetware and Genesee by Shyam Bhakta. <Email for suggestions. Qiagen.com buffers.

Do not autoclave solutions containing ethanol, isopropanol or MOPS; use filter-sterilization if necessary with compatible filter type (nylon for alcohol solns; PES otherwise).
Adjust pH of Tris (base) solutions with HCl.

  • Buffer P1 — miniprep resuspension buffer, store 4°C after RNase addition, cat#19051
    • 50 mM Tris-HCl, pH 8.0
    • 10 mM EDTA
    • 100 mg/L RNase A
  • Buffer P2 — miniprep lysis buffer, cat#19052
    • 200 mM NaOH
    • 1%m/V, 10 g/L SDS
    • 43 mg/L thymophthalein in ethanol (opt)
  • Buffer N3 — miniprep neutralization buffer, cat#19064
    • 4.2 M Gu-HCl, guanidinium chloride
    • 0.9 M KOAc
    • HCl → pH 4.8
  • Buffer PB — DNA-column binding buffer, miniprep opt wash buffer, cat#19066
    • 5 M Gu-HCl
    • 30%V/V isopropanol (300 mL/L)
  • Buffer PE — miniprep final wash buffer, cat#19065
    • 10 mM Tris-HCl, pH 7.5
      5× PE base: 50 mM Tris-HCl pH 7.6
      50× PE base: 0.5 M Tris-HCl pH 7.8 (use 20 mL/L + 180 mL/L water)
    • 80%V/V ethanol (800 mL/L)
  • Buffer EB — miniprep elution buffer
    • 10 mM Tris-HCl, pH 8.5
  • Buffer AE — genomic DNA elution buffer
    • 10 mM Tris-HCl, pH 9.0
    • 0.5 mM EDTA
  • Buffer QG — gel solubilization and column binding buffer, cat#19063
    • 5.5 M GuSCN, guanidinium thiocyanate
    • 20 mM Tris-HCl, pH 6.6
  • Buffer QX1 — gel solubilization and column binding buffer, cat#20912
  • Buffer P3 — QIAfilter/tip DNA prep neutralization buffer
    • 3.0 M KOAc, pH 5.5
      adjust pH with glacial acetic acid, ≈110 mL/L.
  • Buffer DP3 — Qiagen Directprep 96-well miniprep
    • 3.0 M NH₄OAc, pH 5.5
  • Buffer QXB — for binding >3 kb fragments to columns, cat#19054
    • 5 M Gu-HCl
  • Buffer FWB2 — QIAfilter wash buffer
    • 1 M KOAc, pH 5.0
  • Buffer QBT — QIAfilter equilibration buffer
    • 750 mM NaCl
    • 50 mM MOPS, pH 7.0
    • 15% isopropanol (150 mL/L)
    • 0.15%, 1.5 g/L Triton X-100
  • Buffer QC — QIAfilter/tip wash buffer, cat#19055
    • 1.0 M NaCl
    • 50 mM MOPS pH 7.0
    • 15% isopropanol (150 mL/L)
  • Buffer QF — QIAfilter elution buffer, cat#19056
    • 1.25 M NaCl
    • 50 mM Tris-HCl, pH 8.5
    • 15% isopropanol (150 mL/L)
  • Buffer QN  — EndoFree QIAfilter elution buffer
    • 1.6 M NaCl
    • 50 mM MOPS, pH 7.0
    • 15% isopropanol (150 mL/L)
  • Buffer B1 — bacterial lysis buffer
    • 50 mM Tris-HCl, pH 8.0
    • 50 mM EDTA
    • 0.5%, 5 g/L Tween-20
    • 0.5%, 5 g/L Triton X-100
    • 200 μg/L RNase A
  • Buffer B2 — bacterial lysis buffer
    • 3 M Gu-HCl
    • 20%, 200 g/L Tween-20
  • Buffer C1 — cell lysis buffer, store 4°C
    • 1.28 M sucrose
    • 40 mM Tris-HCl, pH 7.5
    • 20 mM MgCl₂
    • 4%, 40 g/L Triton X-100
  • Buffer G2 — digestion buffer
    • 800 mM Gu-HCl
    • 30 mM Tris-HCl, pH 8.0
    • 30 mM EDTA
    • 5%, 50 g/L Tween-20
    • 5%, 50 g/L Triton-X100
  • Buffer Y1 — yeast lysis buffer, store 4̈°C
    • 1 M sorbitol
    • 100 mM EDTA pH 8.0
    • 14 mM β-mercaptoethanol (added just before use)
  • Buffer PAA — PAGE DNA elution
    • 500 mM NH₄OAc
    • 100 mM Mg(OAc)₂
    • 1 mM EDTA
    • 0.1% SDS
  • Buffer TE — DNA elution/resuspension/storage buffer
    • 10 mM Tris-HCl, pH 8.0
    • 0.1 or 1 mM EDTA
  • Buffer STE — DNA resuspension/storage buffer
    • 100 mM NaCl
    • 10 mM Tris-HCl, pH 8.0
    • 1 mM EDTA
  • LyseBlue®, 1000× — pH indicator, clear→blue ≥pH 9.3
    • 43 mg/mL thymophthalein
    • in ethanol
  • Buffer P1 base, 1 L:
    1. 6.06 g Tris base
    2. 2.922 g EDTA
    3. 12 M HCl → pH 8.0, ≈300–400 µL
    • Add 50 µL RNase A to each new 50 mL aliquot, stored 4°C.
  • Buffer P2, 1 L:
    1. 8 g NaOH
    2. 10 g SDS
  • Buffer N3, 1 L:
    1. 401.2 g Gu-HCl, guanidinium chloride
    2. 88.33 g KOAc, potassium acetate
    3. 12 M HCl → pH 4.8, ≈32 mL
  • Buffer PE 50× base (0.5 M Tris-Cl), 0.25 L:
    1. 15.14 Tris base
    2. 12 M HCl → pH 7.8
    3. → 250 mL
  • Buffer PE, 1 L:
    1. 20 mL 50× PE base
    2. 180 mL Milli-Q water
    3. 800 mL Ethanol (pure), → 1 L
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