Agarose gel electrophoresis
Extracted DNAs molecular size is compared with High range DNA marker
A260/A280
1.6-1.9
DNase activity
None detected
Stability
Stable for one year
Description
The overall goal is to isolate chromosomal DNA from leaves. The extraction buffer breaks the cell walls to release the cellular constituents. The buffer contains a detergent cetyl methylammonium bromide (CTAB) that disrupts the cellular membranes to release the DNA, EDTA which minimizes nuclease activity and NaCl that removes polysaccharide. The contaminating RNA is removed by digestion with RNase A. Finally, nucleic acids are precipitated in water-alcohol mixture.
Includes
Extraction buffer - 25 ml
β Mercaptoethanol - 50 mcl
Chloroform - 50 ml
Isoamylalcohol - 2 ml
DNA precipitation solution-50 ml
Wash solution -50 ml
TE Buffer - 2 ml
RNase A (10 mg/ml) – 50 mcl
6X gel loading dye -100 mcl
Agarose -500 mg
Ethidium bromide -30 mcl
50X TAE Buffer -25 ml
DNA marker -60 mcl
