DNA extraction and DNA purification methods

Factors affecting quality of DNA purification:

1. DNase: nuclease contamination in DNA preparation is the major concern for high quality DNA purification. So start with fresh tissue and to process tissue samples quickly is the key to lower the risk of DNA degradation.

2. Shearing of DNA: break down of high molecular weight genomic DNA (and bacterial or yeast artificial chromosomes) by mechnical force should be avoided.

3. Chemical contamination from the DNA purification reagent: contamination of phenol, chloroform all affect downstream application of purified DNA.

DNA purification procedure:

1. Tissue / cell lysis: Don't use too much tissue samples to avoid overloading and shearing of high molecular DNA.

2. DNA extraction procedure to segragate DNA from proteins/chemical contaminatants.

3. DNA precipitation using ethanol, isopropanol, PEG (Polyethylene glycol) or TCA.

4. Wash and dissolving DNA.

Reference:

Molecular biology problem solver: A Laboratory Guide
DNA purification application notes (promega notes)

Last update 18-Oct-2006, Rating n/a of 0 votes.