Fluorescein-High Prime
ROCHE/11585622910 - suitable for dot blot, suitable for hybridization, suitable for Southern blotting, solution
Synonym: DNA labeling
Product Type: Chemical
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 100 μg |
| Quality Level | 100  |
| shipped in | dry ice |
| storage temp. | −20°C |
| technique(s) | dot blot: suitable |
| hybridization: suitable | |
| Southern blotting: suitable | |
| usage | sufficient for 25 labeling reactions (0.01 to 3 ug template DNA per assay) |
| Application: | Fluorescein-High Prime labeled DNA probes has been used in in a variety of hybridization techniques: • in Southern blots • in dot/slot blots • for screening of gene libraries • in in situ hybridizations |
| Components: | 5x concentrated random primer mix: 1U/μL Klenow polymerase, labeling grade, 1mM dATP, 1mM dCTP, 1mM dGTP, 0.65mM dTTP, 0.35mM fluorescein-12-dUTP in 50% (v/v) glycerol. |
| DNA Profile: | DNA fragments of at least 100bp Linearized plasmids, cosmid or λDNA Supercoiled DNA Low level DNA (10ng), e.g., DNA restriction fragments isolated from gels or in molten agarose can be used. |
| General description: | Fluorescein-High Prime is a convenient enzyme and nucleotide mixture for for highly efficient rapid random-primed labeling of DNA with Fluorescein-12-dUTP. 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. Fluorescein-labeled probes are generated at high yield within one hour or after overnight incubation. A standard labeling reaction with 1μg template yields 0.8μg fluorescein-labeled DNA after 1 hour, and 2μg after a 20-hour incubation at +37°C. |
| Other Notes: | Assay Time: 80 minutes |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Principle: | Fluorescein-labeled DNA probes are generated with Fluorescein-High Prime according to the "random primed" labeling technique. The complementary DNA strand is synthesized by Klenow polymerase using the 3′-OH termini of the random oligonucleotides as primers. Fluorescein-12-dUTP is incorporated into the newly synthesized complementary DNA strand. Fluorescein-High Prime is a specially developed reaction mixture containing Fluorescein-12-dUTP and all reagents necessary for random-primed labeling, including Klenow enzyme, premixed in an optimized 5x concentrated reaction buffer concentrate in 50% glycerol. |
| Quality: | Function tested in combination with the DIG RNA Labeling Kit. |
| 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 |