Agarose gel electrophoresis
Extracted DNAs molecular size is compared with High range DNA marker
DNase activity
None detected
A260/A280
1.6-1.9
Stability
Stable for one year
Description
The overall goal is to isolate chromosomal DNA from bacterial cells. The cells are first re-suspended in a suitable buffer (that minimizes nuclease activity) and ruptured to release the cellular contents. The contaminating RNA is removed by digestion with RNase A. Finally, nucleic acid is precipitated in water-alcohol mixture in presence of high concentration of inorganic salt
Includes
Bacterial cell pellet - 18 vials
SE buffer with RNaseA - 5 ml
20% SDS - 200 mcl
2.5 M KCl - 200 mcl
Buffer saturated phenol - 4 ml
Chloroform - 5 ml
DNA precipitation solution - 14 ml
Wash solution - 15 ml
TE buffer - 500 mcl
Agarose - 0.5 g
Ethidium bromide - 30 mcl
6X gel loading dye - 100 mcl
Control DNA (200 ng /10 ml) - 60 mcl
50X TAE Buffer - 25 ml
