Dominik Haudenschild, March 23, 1997

Generating random primed, double stranded labeled probe

• add 25ng DNA template

• bring volume to 25µl with water

• add 10µl random oligo primers

• boil at 95°C for five minutes then quickly spin reaction to bottom of tube

• add 10µl of 5x primer buffer

• add 5µl of labeled nucleotide (3000 to 6000Ci/mmol)

• 1µl Exo(-) Klenow enzyme at 5 U/ml

• mix thoroughly with pipet tip

• incubate 37 to 40°C for 2 to 10 minutes

• add 2µl stop mix

* optional: clean probe over desalting column