1,973
edits
Qiagen Buffers: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
→Formulas for Qiagen Kit Buffers
(New page: ==Formulas for Qiagen Kit Buffers==) |
|||
| Line 1: | Line 1: | ||
==Formulas for Qiagen Kit Buffers== | ==Formulas for Qiagen Kit Buffers== | ||
Do not autoclave solutions containing isopropanol or MOPS; use sterile filtration if necessary. | |||
Buffer P1 | |||
* 50 mM Tris-HCl pH 8.0 | |||
* 10 mM EDTA | |||
* 100 μg/ml RNaseA | |||
The buffer and RNaseA can also be ordered from Qiagen separately (catalog numbers 19051 and 19101). | |||
Buffer P2 | |||
* 200 mM NaOH | |||
* 1% SDS | |||
Buffer P3 (not for spin columns, but for Qiatips, midi, maxi, giga kits) | |||
* 3.0 M potassium acetate pH 5.5 | |||
Buffer N3 | |||
* 4.2 M Gu-HCl | |||
* 0.9 M potassium acetate | |||
* pH 4.8 | |||
Buffer PB | |||
* 5 M Gu-HCl | |||
* 30% ethanol | |||
*(maybe add 10mM Tris-HCL PH 6.6, and that is better) | |||
Buffer PE | |||
* 10 mM Tris-HCl pH 7.5 | |||
* 80% ethanol | |||
Buffer QBT equilibration buffer | |||
* 750 mM NaCl | |||
* 50 mM MOPS pH 7.0 | |||
* 15% isopropanol | |||
* 0.15% triton X-100 | |||
Buffer QC wash buffer | |||
* 1.0M NaCl | |||
* 50 mM MOPS pH 7.0 | |||
* 15% isopropanol | |||
Buffer QF elution buffer | |||
* 1.25M NaCl | |||
* 50 mM Tris-HCl pH 8.5 | |||
* 15% isopropanol | |||
Buffer QN | |||
* 1.6M NaCl | |||
* 50 mM MOPS pH 7.0 | |||
* 15% isopropanol | |||
Buffer FWB2 | |||
* 1M potassium acetate, pH 5.0 | |||
(Source: [http://methodsandreagents.pbwiki.com/], [[media:US6383393.pdf | US Patent 6,383,393]]) | |||
The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9.3 | |||
==Recycling Qiagen Columns== | |||
The blue and purple Qiagen columns are identical in formulation. They are interchangeable and can be reused multiple times after treatment with hydrochloric acid, re-equilibration, and wash. Very likely this protocol can be used with other similar columns. Unused columns can be cheaply purchased in bulk from [http://www.epochbiolabs.com/minispin.asp?pageName=products Epoch Biolabs]. | |||
The reuse protocol is: | |||
* Save the collection tubes and columns after elution of DNA | |||
* Fill the column with 700 μl of 1 M HCl | |||
* Cap and store in an airtight container for at least 24 hours (less than a month) | |||
* Wash the columns and collection tubes in a large beaker of water | |||
* Assemble the column and collection tube and wash the column with 700 μl of DI water, discarding the water | |||
* Repeat | |||
* Fill the column with 700 μl of buffer QBT and spin down, discarding the buffer | |||
* Place the columns in an airtight plastic bag for storage | |||
* Wash the collection tubes, air dry, and store them for reuse | |||
References: PMID 17373483 [http://www.bio.davidson.edu/courses/Molbio/Protocols/QIAQuick_recycle.html Davidson Column Recycling instructions] | |||
