11004760001
Roche
Random Primed DNA Labeling Kit
Synonym(s):
nucleic acid labeling
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About This Item
UNSPSC Code:
41105500
Recommended Products
usage
sufficient for 50 labeling reactions
Quality Level
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.
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.
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.
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.
Kit Components Only
Product No.
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)
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
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Random Primed DNA Labeling Kit Protocol & Troubleshooting
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