High Prime
ROCHE/11585592001 - solution, sufficient for 50 labeling reactions, pkg of 200 μL
Synonym: high prime; labeling mix
Product Type: Chemical
Kinetics of incorporation. 25 ng λDNA was labeled with 50 μCi [α32P]dCTP, 3,000 Ci/mMol.
Kinetics of specific activity. 25 ng λDNA was labeled with 50 μCi [α32P]dCTP, 3,000 Ci/mMol.
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 200 μL |
| Quality Level | 100  |
| storage temp. | −20°C |
| usage | sufficient for 50 labeling reactions |
| Analysis Note: | 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 • DNA fragments of at least 100 bp • Linearized plasmid, cosmid or λDNA • Supercoiled DNA • Or minimal amounts of DNA (10 ng), e.g., DNA restriction fragments isolated from gels or in molten agarose Note: The length of the DNA to be labeled does not influence the reaction. Maximal incorporation may require a prolonged incubation period of 30 to 60 minutes. |
| Application: | 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 • Northern blots • Dot/slot blots • Screening of gene libraries • In situ hybridizations High Prime has been used to label the probes by random hexamer labelling and labelling of probes with 32P by random priming. |
| General description: | 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. |
| General description: | 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. |
| General description: | 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°C. High Prime guarantees efficient labeling of: • DNA amounts ranging from 10 ng to 3 μg in a standard reaction • DNA of different lengths ranging from small restriction fragments to l or cosmid DNA • DNA, supercoiled or linearized • DNA in low melting-point agarose. High Prime-labeled DNA probes are suitable for single-copy gene detection in mammalian DNA in Southern, northern, and dot blots, as well as in colony- or plaque hybridization. |
| General description: | 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. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Principle: | 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. |
| Quality: | Function tested in the standard assay. |
| 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. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | No data available |
| Flash Point(C) | No data available |
| Storage Temp. | −20°C |
| UNSPSC | 41105500 |