Frequently Asked Questions - PurElute™ Bacterial Genomic Kit

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In order to ensure resuspension the pellet must be completely redissolved. Be sure to wash any DNA off the walls, particularly if using V-bottom tubes. It is recommended to use the standard 1.5 ml micro-centrifuge tubes since the pellets can easily collect at the bottom of the tube.

If the eluted sample is too dilute you may re-precipitate your sample by standard alcohol precipitation. (Sambrook J., and Russell D, (eds) (2001) Molecular Cloning: A Lab Manual. 3rd Ed. 6.25, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

Yes. Tris is commonly combined with EDTA for long term storage of DNA. EDTA keeps your DNA stable by chelating heavy metals. It is recommended that genomic DNA be stored in neutral to slightly basic buffered solutions (i.e., 10 mM TrisHCl pH 8.5) to remain more stable.

NOTE: EDTA may inhibit some downstream applications such as restriction enzyme reactions

An elution volume of 100µl is recommended for genomic DNA isolated from cultured cells using the PurElute™ Bacterial Genomic Kit. Smaller elution volumes may lower yield due to insufficient resuspension.

Redissolve by warming in 37° C for a few seconds until the precipitate disappears.

After incubation, the lysate should be clear. A cloudy or turbid lysate indicates an incomplete lysis. This can be due to various reasons including overgrown cells or insufficient mixing during lysis.

Genomic DNA can be accurately quantified by agarose gel electrophoresis, optical density readings at 260nm (OD260), and using Invtrogen Quant-it™ Picogreen® dsDNA Assay Kit.

The PurElute™ Bacterial Genomic Kit produces high-quality genomic DNA. The isolated DNA can be used for agarose gel analysis, restriction enzyme digestion and PCR amplification.

The homebrew method entails purifying genomic DNA from an overnight digestion with proteinase K then extracting with phenol: chloroform followed by alcohol precipitation. Using our PurElute™ Bacterial Genomic Kit is faster, safer and contains no harsh chemicals like phenol and chloroform.

Spheroplast Buffer must be stored at -20ºC and thawed before use. The other components of the PurElute™ Bacterial Genomic Kit should be stored at 4ºC. The kit is stable for up to one year under these conditions.

An elution volume of 100µl is recommended for genomic DNA isolated from cultured cells using the PurElute™ Bacterial Genomic Kit. Smaller elution volumes may lower yield due to insufficient resuspension.

Up to 10 samples can be processed in as little as 15 minutes of hands-on time.

The expected yield of genomic DNA isolated from bacteria with the PurElute™ Bacterial Genomic Kit is up to 40 µg of DNA. However yields of genomic DNA will vary depending on bacterial strain, quality of the starting material, growth conditions, and the amount of material processed.

PurElute™ Bacterial Genomic Kit is intended for use in the isolation of genomic DNA from 2 ml to 5 ml bacterial cultures with an O.D. of 1.0. Rich growth media such as TB (terrific broth) or 2XYT will produce more bacteria (up to 5 times), but this may not lead to greater yields or higher-quality DNA. PurElute™ Bacterial Genomic Kit