Storage: −20°C
UNSPSC Code: 41106000
Components: Control DNA lambda, 20 μl 12.5 ¬µg/ml; dATP, 50 μl 0.5 mM; dCTP, 50 μl 0.5 mM; dGTP, 50 μl 0.5 mM; dTTP, 50 μl 0.5 mM; Hexanucleotide mixture in 10x reaction buffer (100 μl); Klenow enzyme, labeling grade (100 U)
RIDADR: NONH for all modes of transport
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 libraries
• Southern, dot and northern blots
• In situ hybridizations
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.
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.
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.