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Merck

11004760001

Roche

Random Primed DNA Labeling Kit

Synonyme(s) :

nucleic acid labeling

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A propos de cet article

UNSPSC Code:
41105500
NACRES:
NA.54
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usage

sufficient for 50 labeling reactions

manufacturer/tradename

Roche

storage temp.

−20°C

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.

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

Biochem/physiol Actions

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.

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.

Composants de kit seuls

Réf. du produit
Description

  • 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)

Classe de stockage

12 - Non Combustible Liquids

wgk

WGK 1

flash_point_f

does not flash

flash_point_c

does not flash


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Consulter la Bibliothèque de documents

Daniel R Semlow et al.
Cell, 167(2), 498-511 (2016-10-04)
During eukaryotic DNA interstrand cross-link (ICL) repair, cross-links are resolved ("unhooked") by nucleolytic incisions surrounding the lesion. In vertebrates, ICL repair is triggered when replication forks collide with the lesion, leading to FANCI-FANCD2-dependent unhooking and formation of a double-strand break
I J Blader et al.
The Journal of biological chemistry, 276(26), 24223-24231 (2001-04-11)
Cells infected with the intracellular protozoan parasite Toxoplasma gondii undergo up-regulation of pro-inflammatory cytokines, organelle redistribution, and protection from apoptosis. To examine the molecular basis of these and other changes, gene expression profiles of human foreskin fibroblasts infected with Toxoplasma
Juliane Stieber et al.
The Journal of biological chemistry, 280(41), 34635-34643 (2005-07-27)
Hyperpolarization-activated, cyclic nucleotide-gated cation (HCN) channels underlie the inward pacemaker current, termed I(f)/I(h), in a variety of tissues. Many details are known for the HCN subtypes 1, 2, and 4. We now successfully cloned the cDNA for HCN3 from human
Fluorescence in situ hybridization to the polytene chromosomes of anopheles mosquitoes
He F, et al.
PLoS Pathogens (2012)
Margit Dlaska et al.
Cell cycle (Georgetown, Tex.), 12(13), 2084-2099 (2013-06-14)
Immortal cells require a mechanism of telomere length control in order to divide infinitely. One mechanism is telomerase, an enzyme that compensates the loss of telomeric DNA. The second mechanism is the alternative lengthening of telomeres (ALT) pathway. In ALT

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