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Enhanced Avian First Strand Synthesis Kit

Components for cDNA synthesis with enhanced AMV reverse transcriptase

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Synonym(s):
Reverse Transcriptase cDNA synthesis kit, eAMV-RT
NACRES:
NA.55

usage

sufficient for 50 reactions
 kit sufficient for 50 reactions

feature

dNTPs included
hotstart: no

technique(s)

RT-PCR: suitable

color

colorless

input

purified RNA

shipped in

dry ice

storage temp.

−20°C

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This Item
A4464MOD50THIFICDNARO
technique(s)

RT-PCR: suitable

technique(s)

RT-PCR: suitable

technique(s)

-

technique(s)

RT-PCR: suitable, RT-qPCR: suitable

input

purified RNA

input

purified RNA

input

-

input

purified RNA

shipped in

dry ice

shipped in

wet ice

shipped in

-

shipped in

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

20-25°C

storage temp.

-

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

100

General description

Enhanced Avian First Strand Synthesis Kit utilizes a highly purified avian myeloblastosis virus reverse transcriptase (eAMV-RT) that offers superior performance in comparison to standard AMV-RT or standard Moloney murine leukemia virus reverse transcriptase (MMLV-RT). This exceptionally robust eAMV-RT has 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 from total RNA or poly(A)+ RNA.
Sigma′s Enhanced Avian RT First Strand Synthesis Kit also provides random nonamers (9-mers) and anchored oligo (dT)23 primers since gene-specific primers may not always be useful or possible. The anchored oligo has 23 thymidine residues and one G, C, or A residue (the anchor). The anchor ensures that the oligo (dT) primer binds at the very beginning of the message and that there is not a long region of the unusable sequence. With this kit, a dependable cDNA is generated that can be used for various downstream applications, including PCR.

Application

Enhanced Avian First Strand Synthesis Kit has been used for cDNA synthesis, which is intended for use in a real-time reverse transcription-polymerase chain reaction (RT-PCR).

Unit Definition

One unit incorporates one nanomole of TMP into TCA-precipitable material in 10 minutes using polyadenylic acid as template and oligo(dT)12-18 as primer.

Legal Information

Purchase of this product is accompanied by a limited license for use in the Polymerase Chain Reaction (PCR) process for research purposes only and in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by an up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., and authorized thermal cycler.

Kit Components Only

Product No.
Description

  • Enhanced Avian Reverse Transcriptase 1000 U

Kit Components Also Available Separately

Product No.
Description
SDS

  • Deoxynucleotide mix 50 μL

  • O4387Anchored oligo (dT)23 100 μLSDS

  • R7647Random nonamers 100 μLSDS

  • O4387Ribonuclease inhibitor 50 μLSDS

  • W1754PCR grade water 1.5 mLSDS

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Articles

One approach to the analysis of gene expression is to measure the concentration of mRNA of a gene. There are several challenges to such analyses, such as the differences in half life between different transcripts, the temporal patterns of transcription and the lack of correlation between mRNA and protein.

Protocols

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 degradation is less than that detected by capillary systems but still sufficient to effect qPCR analyses.

Related Content

Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.

RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), and detecting pathogens such as viruses.

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