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Invitrogen™ RPA III™ Ribonuclease Protection Assay Kit

Fast single-tube reaction

£195.00 - £476.45

Specifications

Storage Requirements Store Hybridization III Buffer, RNase Digestion III Buffer, Yeast RNA, RNase Inactivation/Precipitation III Solution, Gel Loading Buffer II, pTRI-Actin-Mouse, RNase A/RNase T1 Mix, RNase T1, and 5 M Ammonium Acetate at -20°C. Store Mouse Liver RNA at below -70°C. Store Probe Elution Buffer at 4°C.
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Includes: Hybridization III Buffer, RNase Digestion III Buffer, Yeast RNA, RNase Inactivation/Precipitation III Solution, Gel Loading Buffer II, pTRI-Actin-Mouse, RNase A/RNase T1 Mix, RNase T1, 5M Ammonium Acetate, Mouse Liver RNA, Probe Elution Buffer

Products 3
Product Code Brand Includes Quantity Price Quantity & Availability  
Product Code Brand Includes Quantity Price Quantity & Availability  
10117304 View Documents Invitrogen™
AM1415
Kit without manual 35 reactions £195.00
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Estimated Shipment date
from Supplier 23-12-2019
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10690026 View Documents Invitrogen™
AM1414M
Kit with manual 100 reactions This item has been discontinued and is no longer available. View the product for possible alternatives or contact our Product Support team on 01509 555888 for assistance
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10554635 View Documents Invitrogen™
AM1414
Kit without manual 100 reactions This item has been discontinued and is no longer available. View the product for possible alternatives or contact our Product Support team on 01509 555888 for assistance
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Description

Description

The Ambion kit contains reagents for performing ribonuclease protection assays, an extremely sensitive technique for the detection, quantitation, and characterization of RNA.

  • Ideal for new users
  • Simplified single-tube reaction
  • Sensitive: detect as little as 5fg of target mRNA
  • No phenol extraction or Proteinase K digestion
  • Ideal for multiprobe-analysis: up to 12 genes analyzed simultaneously in each sample
  • Used for detection, quantitation, and mapping of mRNA

Fast, Sensitive Assay
The procedure for the RPA III Kit is fast and takes place in a single microfuge tube. Hybridization occurs in solution rather than on a solid support (i.e., nylon membrane), allowing more complete hybridization of the antisense probe to the target mRNA. After hybridization, excess single-stranded probe and unhybridized sample RNA are removed by digestion with a mixture of ribonucleases. Then, in a single step, ribonucleases are inactivated, and the protected double-stranded RNA (probe + target) is precipitated. The products are separated on a denaturing polyacrylamide gel and then visualized by autoradiography when isotopic probes are used or transferred to a membrane for nonisotopic detection protocols.

Use Multiple Probes with a Single RNA Sample
In current research facilities, it is rarely sufficient to assess the expression level of a single gene. Ideally, multiple targets and one or two internal controls would be assessed within individual samples. For such analyses, nuclease protection assays are without peer among the commonly employed methods of RNA analysis. Five to ten probes plus one or two internal controls can routinely be used in a single assay, allowing simultaneous quantitation of multiple messages within a single RNA sample.

Additional Benefits of RPAs
Northern blots are limited by the amount of total RNA that can be loaded into a single lane without overloading the gel (typically approx.20μg per lane). With ribonuclease protection assays as much as 100μg of RNA may be used since hybridization takes place in solution, and the nonhybridized RNA will be degraded before gel analysis. Additionally, because probes for protection assays are significantly shorter than the mRNA species being detected, the target RNA need not be completely intact (breaks that occur outside the region that hybridizes to the probe will not affect RPA data but will result in smeared bands on northerns).

DNA and RNA Purification and Analysis, Nuclease Protection Assays, Nucleic Acid Gel Electrophoresis and Blotting

Order Info

Shipping Condition: Dry Ice

Specifications

Specifications

Store Hybridization III Buffer, RNase Digestion III Buffer, Yeast RNA, RNase Inactivation/Precipitation III Solution, Gel Loading Buffer II, pTRI-Actin-Mouse, RNase A/RNase T1 Mix, RNase T1, and 5 M Ammonium Acetate at -20°C. Store Mouse Liver RNA at below -70°C. Store Probe Elution Buffer at 4°C.

For Research Use Only. Not for use in diagnostic procedures.