Not available outside of the UK & Ireland.

Application

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 librariesSouthern, dot and northern blotsIn situ hybridizations

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

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.

Other Notes

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

Packaging

1 kit containing 7 components.

Preparation Note

Working solution: Preparation dNTP Stock MixTo 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 dATP1 µl dGTP1 µl dTTP to a reaction vialPreparation of DIG Stock MixTo 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.

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.