Recommended Methods of Purification
The mere use of the kits we recommend does not guarantee success.
- Qiagen Spin Kits- Note: Always do the optional wash even if your strain is not part of the JM series. The elution buffer that comes with the kit works well for sequencing. No need to use dH2O.
Qiagen publishes a very good template purification guide. Please ignore its advice on genomic preparation as we do not accept genomic preps. If you are having trouble getting yield with the Spin kits: Elute with hot dH2O (60C-70C) that has been recently autoclaved.
- Promega Wizard SV
- Bio-Rad Plasmid Miniprep Kit
- Nucleobond AX Cartridge
- Novagen UltraMobius 200 Plasmid Kit
- Mini-alkaline lysis + PEG precipitation Note: This is labor-intensive, requires some skill, and nobody uses it anymore. You’re better off purchasing a kit.
Purification Methods Not Recommended
- Any Qiagen MIDI or MAXI kit! Some people can make these work for sequencing but we do NOT recommend them. They have proven to be unreliable and inconsistent for sequencing.
- S & S DNA Elution Beads
- Stratagene Strataresin
- Isoquick Plasmid Kit
- Classical Alkaline Lysis
- 3′-5′ Plasmid Isolation Kit. Although Perkin Elmer recommends it, it seems to be very inconsistent.
- Gibco’s Concert Prep.
- BIO 101 Genekleen
- CsCl purification
PCR Purification Methods
- QIAquick Gel Extraction Kit. (cat#28704)
- QIAquick PCR Purification Kit (cat#28104) This will NOT work for sequencing if you have multiple bands! There are also problems with getting rid of excess primers. Optimize your PCR reaction!
- Wizard® SV Gel and PCR Clean-Up System
- Zymoclean Gel DNA Recovery kit ; (Protocol shared by Gina DiGregorio from Phillip Kern’s laboratory)
Online Analysis Tools
Disclaimer: We do not provide technical support for the following links nor are we responsible for the results obtained by using them. We are merely making known to you the free computing resources that are available to the public.
- National Center For Biotechnology Information
- NCBI Online Tools
- Molecular Biology Freeware For Windows
- Molecular Biology Freeware for Macintosh
- Protein Information Resource
- Swiss Institute of Bioinformatics Tools