Sunteți pe pagina 1din 2

Qiagen Plasmid Prep - Buffer Composition

http://people.mbi.ucla.edu/sumchan/qiagenbuffer.html

Qiagen Plasmid Prep - Buffer Compositon and Preparation


Note: Buffer compositions are given per liter of solution. Do not autoclave isopropanol-containing buffer, but sterilize by filtration instead. Buffer calculations are base on Tris base adjusted to pH with HCl (Tris-Cl). If using Tris-HCl reagent, the qualities used should be recalculated. Buffer P1 - Resuspension Buffer 50mM Tris-Cl, pH 8.0, 10mM EDTA, 100ug/mL RNase A Storage condition - 4 C after adding RNase A Prep - Dissolve 6.06g Tris base, 3.72g EDTA-2H 0 in 800mL dH20. Adjust the pH to 8.0 with HCl.
2 o

Adjust the volume to 1 liter with dH O. Add 100mg RNase A per liter of P1.
2

Buffer P2 - Lysis Buffer 200mM NaOH, 1% SDS Storage condition - RT Dissolve 8.09g of NaOH pellets in 950mL dH O, 50mL 20% SDS solution. The final volume should
2

be 1 liter. Buffer P3 - Neutralization Buffer for Qiatips, Midiprep, Maxiprep, and Gigaprep kits. 3.0M potassium acetate, pH 5.5 Storage condition - 4 C or RT Dissolve 294.5g potassium acetate in 500mL dH O. Adjust the pH to 5.5 with glacial acetic acid
2 o

(about 110mL). Adjust the volume to 1 liter with dH O.


2

Buffer N3 - Neutralization Buffer for spin columns. Composition unknown Storage condition - RT Buffer PE - Wash Buffer Composition unknown Storage condition - RT Buffer QBT - Equilibrium Buffer 750mM NaCl, 50mM MOPS, pH7.0, 15% isopropanol, 0.15% Triton X-100 Storage condition - RT Dissolve 43.83g NaCl, 10.46g MOPS (free acid) in 800mL dH O. Adjust the pH to 7.0. Add 150mL
2

pure isopropanol and 15mL 10% Triton X-100 solution. Adjust the volume to 1 liter with dH O.
2

Buffer QC - Wash Buffer 1.0M NaCl, 50mM MOPS, pH 7.0, 15% isopropanol Storage condition - RT Dissolve 58.44g NaCl and 10.46g MOPS (free aicd) in 800mL dH O. Adjust the pH to 7.0. Add
2

150mL pure isopropanol. Adjust the volume to 1 liter with dH O.


2

Buffer QF - Elution Buffer 1.25M NaCl, 50mM Tris-Cl, pH 8.5, 15% isopropanol

1 of 2

2/21/2013 4:50 PM

Qiagen Plasmid Prep - Buffer Composition

http://people.mbi.ucla.edu/sumchan/qiagenbuffer.html

Storage condition - RT Dissolve 73.05g NaCl and 6.06g Tris base in 800mL dH O and adjust the pH to 8.5 with HCl. Add
2

150mL pure isopropanol. Adjust the volume to 1 liter with dH O.


2

TE Buffer 10mM Tris-Cl, pH 8.0, 1mM EDTA Storage condition - RT STE Buffer 100mM NaCl, 10mM Tris-Cl, pH 8.0, 1M EDTA Storage condition - RT Dissolve 5.84g NaCl, 1.21g Tris base and 0.37g EDTA-2H O in 800mL dH O and adjust the pH to
2 2

8.0 with HCl. Add up the volume to 1 liter with dH O.


2

EB Buffer - Elution Buffer 10mM Tris-Cl, pH 8.5 Storage condition - RT QG Buffer - Solubilization Buffer for Gel Extration Kit Composition Unknown (Proprietary) Storage condition - RT PB Buffer - Binding Buffer Composition Unknown (Proprietary) Storage condition - RT Return to Experimental Procedures and Solution Recipes Return to Home

2 of 2

2/21/2013 4:50 PM