产品说明
一般描述
High Prime, a novel labeling mixture, contains optimal concentrations of nucleotides and primers in a highly efficient reaction mix with Klenow enzyme. This convenient "all-in-one" principle of High Prime reduces pipetting steps to a minimum, and increases accuracy and reproducibility of labeling reactions.
Enzyme and nucleotide mixture for rapid random-primed labeling of DNA with [32P], [35S], or [3H] dCTP. 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.
Contents:
5x concentrated random primer mix: 1 U/μl Klenow polymerase, labeling grade, 0.125 mM dATP, 0.125 mM dGTP, 0.125 mM dTTP in 50% (v/v) glycerol.
High Prime is used for rapid random-primed labeling of DNA with [32P], [35S], or [3H]dCTP to a guaranteed specific activity of >2 x 109 dpm/μg. Incubation time is only 10 minutes at +37℃.
High Prime guarantees efficient labeling of:
特异性
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0) and/or by heating to 65 ℃ for 10 minutes.
应用
For the labelling of DNA with radioactive dCTP using random oligonucleotides as primers.
High Prime-labelled probes are used in a variety of hybridization techniques: Southern blots
质量
Function tested in the standard assay.
原理
Labeled probes are generated with High Prime according to the random-primed labeling technique. High Prime is a specifically developed reaction mixture containing random oligonucleotides, Klenow polymerase, labeling grade, dATP, dGTP, and dTTP in an optimized reaction buffer concentrate in 50% glycerol for rapid and efficient labeling of DNA with [32P]-, [35S]-, or [3H]-labeled dCTP.
分析说明
Specific Activity: The standard assay routinely yields a specific activity of 2 x 109 dpm/μg, using different substrate DNAs after 10 minutes of incubation.
Assay Time: 30 minutes
Sample Materials
其他说明
仅用于生命科学研究。不可用于诊断。
产品性质
形式 | solution |
质量水平 | 100 |
用途 | sufficient for 50 labeling reactions |
包装 | pkg of 200 μL |
manufacturer/tradename | Roche |
运输 | dry ice |
储存温度 | −20℃ |
安全信息
储存分类代码 | 12 - Non Combustible Liquids |
WGK | WGK 1 |
闪点(F) | No data available |
闪点(C) | No data available |
Sigma-Aldrich