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Invitrogen™ KinaseMax™ 5' End-Labeling Kit
Description
Includes
T4 Polynucleotide Kinase, Calf Intestine Alkaline Phosphatase, 10X Kinase Buffer, Dephosphorylation Buffer, Phosphatase Removal Reagent, RNA Transcript Control, pUC19/Sau3 A Markers, ATP Solution, Gel Loading Buffer II, Nuclease-free water
The Invitrogen KinaseMax™ 5' End-Labeling Kit allows the efficient end-labeling of DNA or RNA to high-specific activity with T4 polynucleotide kinase and [gamma-32P] ATP, or quantitative phosphorylation of 5' ends using unlabeled ATP.
The kit includes a kinase reaction buffer that exceeds the performance of the standard forward reaction buffers recommended in “Molecular Cloning: A Laboratory Manual” (Sambrook et al., 1989) and “Current Protocols in Molecular Biology” (John Wiley & Sons, Inc. Ausubel, FM et al. editors, 1994). Crude 7000 Ci/mmol [gamma-32P]ATP is compatible with this kit. End-labeled probes are more stable than internally labeled probes and can routinely be used in assays for two to four weeks.
Rapid, Phenol-Free Phosphatase Removal Step
Inactivating the Calf Intestinal Phosphatase (CIP) after the dephosphorylation reaction is a time-consuming and cumbersome procedure that usually requires phenol extraction and ethanol precipitation typically resulting in sample loss. The KinaseMax Kit now includes a novel Phosphatase Removal Reagent for quick and complete removal of CIP. After the dephosphorylation step, the reagent is added to the reaction, incubated at room temperature for 2 minutes. The tube is then spun for a minute in the microfuge and the supernatant transferred to a new tube for the kinasing reaction.
Accessory Products:
The NucAway™ Spin Columns (Cat. No. AM10070) are available for rapid purification of end-labeled probes.
- Includes sufficient reagents for 30 reactions
- Label oligonucleotide probes for nuclease protection assays and blot hybridizations
- Label primers for northerns and quantitative PCR
- Obtain up to 3-fold greater yields than with standard kinase buffers
- Reagents included for forward and dephosphorylation reactions
DNA and RNA Purification and Analysis, DNA Labeling, Northern Blotting, Nuclease Protection Assays, Nucleic Acid Gel Electrophoresis and Blotting, Nucleic Acid Labeling and Oligo Synthesis, Oligo Labeling, RNA Labeling, Southern Blotting
Order Info
Shipping Condition: Dry Ice
Specifications
Specifications
| Product Type | Labeling Kit |
| Detection Method | Radioactive |
| Assay | Nuclease Protection Assay |
| Content And Storage | T4 Polynucleotide Kinase, Calf Intestine Alkaline Phosphatase, 10X Kinase Buffer, Dephosphorylation Buffer, Phosphatase Removal Reagent, RNA Transcript Control, pUC19⁄Sau3 A Markers, ATP Solution, Gel Loading Buffer II should be stored at –20°C. Nuclease-free water may be stored at any temperature. |
| Final Product Type | Probes (Labeled DNA), Probes (Labeled RNA) |
| Label or Dye | 32P (Phosphorous-32) |
| Includes Label or Dye | No |
| Labeling Target | cDNA, DNA (General), PCR Products, Oligos, RNA (General) |
| Labeling Method | Direct Labeling |
| Shipping Condition | Dry Ice |
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For Research Use Only. Not for use in diagnostic procedures.
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