Setting up an analytical restriction digest

Setting up an analytical restriction digest.

Download and Excel file to automatically calculate  Master-mix volumes here. You could run your digests on slide agarose gels or standard agarose gels to resolve the DNA species. Slide agarose gels are faster and are my desired gel system for all DNA and RNA applications.

E. coli transformation and plating

Transforming E. coli with plasmids and plating the transformation.

I have found that the Inoue, H., Nojima, H., & Okayama, H. (1990). Gene, 96(1), 23-28. method gives the highest efficiency competent cells and is very easy to do. You can get a protocol for the Inoue method here.

Plant genomic DNA extraction

The CTAB method yields high quality plant genomic DNA that can be used for virtually any application including Illumina sequencing. The method is fast, cheap, and scalable. For genotyping, less than 1 mm x 1 mm pieces of plant material can be use.

Download a printable CTAB plant genomic DNA extraction protocol here.