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Team:TU Darmstadt/Materials/Quiagen buffers
From 2012.igem.org
Qiagen Kit Buffers
Do not autoclave solutions containing ethanol, isopropanol or MOPS; use sterile filtration if necessary.
Buffer AE (elution buffer for genomic DNA preps)
- 10 mM Tris-HCl
- 0.5 mM EDTA
- pH 9.0
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 DP3 (for Qiagen Directprep 96-well miniprep)
- 3.0 M ammonium 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% isopropanol
Buffer PE
- 10 mM Tris-HCl pH 7.5
- 80% ethanol
Buffer QX1 (for solution and binding of agarose gels)
- 7 M NaPO4
- 10 mM NaAc
- pH 5.3
Buffer QXB (for binding of large >3000 bp fragments to columns)
- 5 M GuHCl
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
Buffer B1 (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
Buffer B2 (bacterial lysis buffer)
- 3 M Gu-HCl
- 20% Tween-20
Buffer C1 (cell lysis buffer) (store at +4°C)
- 1.28 M sucrose
- 40 mM Tris-HCl pH 7.5
- 20 mM MgCl2
- 4% Triton X-100
Buffer G2 (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
Buffer Y1 (yeast lysis buffer) (store at +4°C)
- 1 M Sorbitol
- 100 mM EDTA pH 8.0
- 14 mM beta mercaptoethanol (added just before use)
Buffer PAA (PAGE gel elution of DNA)
- 500 mM NH4Ac
- 100 mM MgAc2
- 1 mM EDTA
- 0.1% SDS
The LyseBlue indicator dye added to some of the buffers is Thymophthalein, pH shift from colorless to blue at pH 9.3
References
[1] http://methodsandreagents.pbwiki.com/
[2] [http://openwetware.org/images/1/1f/US6383393.pdf US Patent 6,383,393]
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
[1] http://www.pubget.com/paper/17373483
[2] http://www.bio.davidson.edu/courses/Molbio/Protocols/QIAQuick_recycle.html
