Economic substitute for Qiagen columns and kits with solution recipes

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  • Joe Phillips 18:37, 25 October 2013 (EDT):Here is our lab note on how to save money on DNA and RNA kits by using home made solutions (or buy bulk solutions)and bulk columns to assemble our own RNA and DNA kits.

Solution Recipes for Qiagen Kits:

NOTE:

•For long tern storage, all the buffers should be sterilized by filtration or autoclaving. Solutions that contain ethanol, isopropanol or MOPS should be sterilized by filtration only. •You can also buy bulk solutions from Qiagen (Cat# is listed below) •The Blue and Purple Qiagen columns are identical. You can reuse the columns after treating with 1M HC, wash with ddH2O, re-equilibrate with buffer QBT, and spin to dry the columns. Economic substitute for Qiagen columns can be purchased in bulk from Enzymax (www.enzymax.net[1]). Qiagen MinElute colums for small samples can also be replaced by bulk Tini Spin Columns from Enzymax (both DNA and RNA columns are available).

Solutions:

P1 (resuspension buffer): (Qiagen Cat# 19051, 500ml) 50 mM Tris-HCl, 10 mM EDTA, pH 8.0 (25oC), 50-100 ug/ml RNase A (Qiagen cat# 19101)

P2 (lysis buffer): (Qiagen Cat#19052, 500ml) 200 mM NaOH, 1% SDS

N3 (neutralization buffer for DNA binding to silica membrane colums): (Qiagen Cat#19064, 500 ml) 4.2 M guanidine hydrochloride (GuHCl), 0.9 M potassium acetate, pH 4.8

P3 (neutralization buffer for Qiatips, midi, giga kits, DO NOT USE THIS for spin columns, use N3 instead) 3.0 M potassium acetate, pH 5.5

DP3 (neutralization buffer for Qiagen Directprep 96-well miniprep) 3.0 M ammonium acetate, pH 5.5

PB Buffer (extra wash step for EndA+ strains*or PCR kit): (Qiagen Cat#19066, 500 ml) 5 M guanidine hydrochloride (Gu-HCL) 30% isopropanol

5x PE Buffer (add ethanol to 80% before use): (Qiagen Cat#19065, 100ml for making 500ml 1x PE Buffer) 80 mM NaCl, 8 mM Tris-HCl, pH 7.5 (25oC)

EB Buffer (DNA elution): 10 mM Tris-HCl, pH 8.0 or ddH2O.

QG Buffer (gel solubilization buffer): (Qiagen cat# 19063, 250 ml) 5.5 M guanidine thiocyanate (GuSCN), 20 mM Tris-HCl, pH 6.6 (25oC), dissolve in pH 7 standard solution (yellow) or water.

AE (elution buffer for genomic DNA isolation): 10 mM Tris-HCl, pH 8.0 0.5 mM EDTA, pH9.0

QX1 (solubilization and binding buffer for agarose gels): (Qiagen cat#20912, 500ml) 7M NaPO4 10mM NaAc, pH 5.3

QXB (for binding of DNA fragments >3.0kb to spin columns): 5M GuHCl

PAA (PAGE gel elution for DNA): 500 mM NH4Ac 100 mM MgAc2 1 mM EDTA 0.1% SDS

QBT (Equilibration buffer): (Qiagen cat# 19054, 1L) 750 mM NaCl 50mM MOPS, pH 7.0 15% Isopropanol 0.15% Triton X-100

QC (Wash buffer): (Qiagen cat# 19055, 1L) 1.0M NaCl 50mM MOPS, pH7.0 15% isopropanol

QF (Elution buffer): (Qiagen cat# 19056, 1L) 1.25M NaCl 50mM Tris-HCl, pH 8.5 15% isopropanol

QN Buffer: 1.6M NaCl 50mM MOPS, pH7.0 15% isopropanol

FWB2 Buffer: (QIAfilter wash buffer) 1M Potassium acetate, pH 5.0

B1 buffer (Bacterial lysis buffer): 50 mM Tris-HCl pH 8.0 50 mM EDTA pH 8.0 0.5% Tween-20 0.5% Triton-X100 RNAse A 200 μg/l

B2 buffer (Bacterial lysis buffer): 3M GuHCl 20% Tween-20

C1 buffer (Cell lysis buffer): 40C storage 1.28 M sucrose 40 mM Tris-HCl pH 7.5 20 mM MgCl2 4% Triton X-100

G2 buffer (Digestion buffer): 800 mM GU-HCl 30 mM Tris-HCl pH 8.0 30 mM EDTA pH 8.0 5% Tween-20 5% Triton-X100

Y1 (Yeast lysis Buffer): 40C storage 1 M Sorbitol 100 mM EDTA pH 8.0 14 mM beta mercaptoethanol (add before use)

LyseBlue (pH indicator dye) 1000x: pH shift from colorless to blue at pH9.3 43 mg/ml Thymophthalein in 100% ethanol.

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