Qiagen Plasmid Prep - Buffer Compositon and Preparation

Note: Buffer compositions are given per liter of solution. Do not autoclave isopropanol-containingbuffer, but sterilize by filtration instead. Buffer calculations are base on Tris base adjusted to pHwith HCl (Tris-Cl). If using Tris-HCl reagent, the qualities used should be recalculated.

Buffer P1 - Resuspension Buffer50mM Tris-Cl, pH 8.0, 10mM EDTA, 100ug/mL RNase AStorage condition - 4oC after adding RNase APrep - 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 per liter of P1.

Buffer P2 - Lysis Buffer200mM NaOH, 1% SDSStorage condition - RTDissolve 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 5.5Storage condition - 4oC or RTDissolve 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.Composition unknownStorage condition - RT

Buffer PE - Wash BufferComposition unknownStorage condition - RT

Buffer QBT - Equilibrium Buffer750mM NaCl, 50mM MOPS, pH7.0, 15% isopropanol, 0.15% Triton X-100Storage condition - RTDissolve 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 Buffer1.0M NaCl, 50mM MOPS, pH 7.0, 15% isopropanolStorage condition - RTDissolve 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 Buffer1.25M NaCl, 50mM Tris-Cl, pH 8.5, 15% isopropanol

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Storage condition - RTDissolve 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.

TE Buffer10mM Tris-Cl, pH 8.0, 1mM EDTAStorage condition - RT

STE Buffer100mM NaCl, 10mM Tris-Cl, pH 8.0, 1M EDTAStorage condition - RTDissolve 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 Buffer10mM Tris-Cl, pH 8.5Storage condition - RT

QG Buffer - Solubilization Buffer for Gel Extration KitComposition Unknown (Proprietary)Storage condition - RT

PB Buffer - Binding BufferComposition Unknown (Proprietary)Storage condition - RT

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