RANDOM PRIMED DNA LABELING KIT

General description

Random primed DNA labeling kit is used for the radioactive and nonradioactive labeling of DNA with modified deoxyribonucleoside triphosphate using random oligonucleotides as primers. In this method, the complementary DNA strand of denatured DNA is synthesized by Klenow polymerase using the 3′-OH termini of the random oligonucleotides as primers.

Specificity

Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0) and/or by heating to 65 °C for 10 minutes.

Application

Random Primed DNA Labeling Kit has been used to label probe or fragments shorter than 1 kb.

Random Primed DNA Labeling Kit is used to uniformly label plasmid or phage DNA with any [α-32P]-dNTP or modified dNTP. Labeled DNA probes with high specific activity are used in a variety of hybridization techniques including:

• Screening of gene libraries
• Southern, dot and northern blots
• In situ hybridizations

Packaging

1 kit containing 7 components.

Preparation Note

Working solution: Preparation dNTP Stock Mix
To avoid pipetting inaccuracy, due to low volumes, prepare a stock mix of unlabeled dNTPs. Aliquots should be stored at -15 to -25 °C. Avoid repeated freezing and thawing.

• dATP, dGTP, dTTP mixture:
  For one labeling reaction pipette:
  1 μl dATP
  1 μl dGTP
  1 μl dTTP to a reaction vial

Preparation of DIG Stock Mix
To avoid pipetting inaccuracy due to low volumes, prepare a DIG stock mix. To do this, mix digoxigenin-11-dUTP [3 mM] and dTTP (vial 5) 1:1. For each labeling reaction 1.6 μl are used.

Other Notes

For life science research only. Not for use in diagnostic procedures.