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HSRT100 Sigma-Aldrich

Enhanced Avian HS RT-PCR Kit

Flexible kit for one-step or two-step RT-PCR

  •  NACRES NA.55

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Properties

Related Categories Molecular Biology, PCR/Amplification, RT-PCR and RT-qPCR More...
Quality Level   200
feature   hotstart
color   colorless
shipped in   wet ice
storage temp.   −20°C

Description

Other Notes

Reverse Transcriptase PCR (RT-PCR) is a powerful tool used to study gene expression. The Enhanced Avian HS RT-PCR kit utilizes an enhanced avian myeloblastosis virus reverse transcriptase (eAMV-RT) enzyme that offers superior performance in comparison to standard AMV-RT and Moloney murine leukemia virus reverse transcriptase (MMLV-RT). eAMV RT is an exceptionally robust enzyme with an enhanced ability to transcribe through difficult secondary structure at elevated temperatures (up to 65 °C) making it the ideal enzyme for producing high quality full-length cDNA (up to 14.1 kb) from total RNA or poly(A)+ RNA. JumpStart AccuTaq LA DNA polymerase mix is also provided to eliminate non-specific amplification and increase specificity and sensitivity. The combination of these two enzymes provides a quality system that offers the versatility of a one-step or two-step RT-PCR protocol.

Application

Enhanced Avian HS RT-PCR Kit has been used to synthesize the cDNA first-strand during reverse transcriptase PCR (RT-PCR) analysis.

Procedures are provided for one-step and two-step RT-PCR reactions.

One-step: In a single tube, eAMV RT and AccuTaq LA act sequentially to first produce cDNA and then immediately amplify by PCR. This provides quick, sensitive analysis of RNA.

Two-step: Each reaction is individually optimized for greater yields with high fidelity, when protocol requires multiple amplifications, or if maximum yield is more important than maximum convenience.

Features and Benefits

• Greater transcription lengths than other reverse transcriptases, generating cDNA up to 14.1 Kb.
• Higher sensitivity for detecting low abundance messages. eAMV RT is able to transcribe RNA that other reverse transcriptases cannot detect.
• Unsurpassed transcription through difficult secondary structure.
• Increased sensitivity, specificity and yield from JumpStart AccuTaq LA DNA Polymerase.

Legal Information

No license is conveyed with the purchase of this product under any of US Patents Nos. 5,804,375, 5,994,056, 6,171,785, 6,214,979, 5,538,848, 5,723,591, 5,876,930, 6,030,787, and 6,258,569, and corresponding patents outside the United States, or any other patents or patent applications, relating to the 5′ Nuclease and dsDNA-Binding Dye Processes. For further information contact the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

AccuTaq is a trademark of Sigma-Aldrich Co. LLC

JumpStart is a trademark of Sigma-Aldrich Co. LLC

eAMV is a trademark of Sigma-Aldrich Co. LLC

Kit component also available separately

Description

Product #

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Anchored oligo(dT)23 primers 100 μL SDS O4387
Anchored oligo(dT)23 primers 100 μL SDS O4387
10x reaction buffers 10x AccuTaq buffer SDS B0174
10 mM dNTP mix 10 x PCR buffer SDS P2192
Nuclease-free water 4 x 1.5 mL SDS W1754
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable
Protocols & Articles

Articles

Reverse Transcription - PCR Technologies Guide

Use the table of contents on the right to navigate to other sections of A Technical Guide to PCR Technologies.
A Technical Guide to PCR Technologies
Keywords: Degradations, Gene expression, Nucleic acid hybridization, PAGE, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Transcription

Protocols

Long and Accurate PCR Amplification of DNA with RedAccuTaq D4812 Protocol

Reaction Optimization Reliable amplification of long DNA sequences requires: 1) effective denaturation of DNA template, 2) adequate extension times to produce large products and 3) protection of targ...
Keywords: AGE, Amplification, Buffers, Diagnostic, Electrophoresis, Evaporation, Gas chromatography, Gel electrophoresis, Melting, Nucleic acid annealing, Nucleic acid denaturation, Phase transitions, Polymerase chain reaction, Size-exclusion chromatography, Solvents

The 3'/5' Assay for Analysis of RNA Integrity Protocol

The 3’/5’ integrity assay is a potential first step in the identification of RNA degradation. The assay is particularly useful when a large number of samples are to be analyzed or when the degradatio...
Keywords: Amplification, Degradations, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

Related Content

PCR Selection Guide

We offer a wide variety of PCR enzymes, master mixes, and PCR protocols to meet your experimental needs for routine PCR, qPCR, or RT-PCR. Our PCR Selection Guide features various filters to sort by, ...
Keywords: Genomics, Polymerase chain reaction, Polymerase chain reaction - quantitative

RT-PCR | Reverse Transcription PCR

Reverse transcription polymerase chain reaction (RT-PCR) is a variation of standard PCR that involves the amplification of specific mRNA obtained from small samples. It eliminates the need for the te...
Keywords: Amplification, Cloning, Diagnostic, Gas chromatography, Molecular biology, Nucleic acid hybridization, PAGE, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Transcription

Peer-Reviewed Papers
15

References

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