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A4464

Sigma-Aldrich

Enhanced Avian Reverse Transcriptase [eAMV™ RT]

For reverse transcription at higher temperatures & rare mRNAs

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About This Item

MDL number:
MFCD01634820
UNSPSC Code:
12352204
NACRES:
NA.55

Quality Level

200

form

crystalline powder

usage

sufficient for 50 reactions

feature

dNTPs included: no
hotstart: no

concentration

20 units/μL

technique(s)

RT-PCR: suitable

color

colorless

input

purified RNA

shipped in

wet ice

storage temp.

−20°C

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General description

eAMV™ Reverse Transcriptase is an enhanced form of Avian Myeloblastosis Virus (AMV) RT that synthesizes a DNA strand complementary to RNA, DNA, or an RNA:DNA hybrid. This exceptionally robust AMV RT has greater thermostability than standard AMV or Moloney murine leukemia virus(M-MLV) reverse transcriptase. eAMV™ RT is an ideal enzyme for producing high-quality full-length cDNA from total RNA or poly(A)+ RNA and is also efficient at transcribing long targets.

Application

Enhanced Avian Reverse Transcriptase (eAMV RT) is used to transcribe RNA into DNA, and facilitates efficient mRNA template driven sysnthesis of cDNAs. This is due to the abillity of this enhanced AMV-RT to transcribe large mRNA templates, to transcribe through difficult secondary structures, and to detect low abundance mRNAs by RT-PCR.
Enhanced Avian Reverse Transcriptase [eAMV™ RT] has been used for reverse transcription of total RNA to synthesize cDNA during quantitative reverse transcription PCR (RT-qPCR) analysis.

Biochem/physiol Actions

Reverse transcriptase catalyzes RNA template incorporation of dNTPs into complimantary DNA through phosphodiester bond formation.

Features and Benefits

  • Greater sensitivity for low abundance mRNA
  • Unsurpassed transcription through difficult secondary structures at elevated temperatures (up to 65°C)
  • Efficient generation of full-length cDNA, up to 14.1 kb
  • Produces first strand cDNA ready for PCR amplification

Packaging

Provided with a vial of 10× reaction buffer.

Unit Definition

One unit incorporates one nanomole of TMP into TCA precipitable material in 10 min using polyadenylic acid as template and oligo(dT)12-18 as a 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.
eAMV is a trademark of Sigma-Aldrich Co. LLC

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 5-5 (1989)
C Tosh et al.
Acta virologica, 41(3), 153-155 (1997-06-01)
A method of reverse transcription (RT) and polymerase chain reaction (PCR) amplification of 1D (VP1) gene of foot-and-mouth disease (FMD) virus using one reaction mixture containing both avian myeloblastosis virus (AMV) reverse transcriptase (RTase) and Tfl DNA polymerase is described.
Emily Schifano et al.
Virulence, 10(1), 1013-1025 (2019-11-28)
Calcium signaling can elicit different pathways involved in an extreme variety of biological processes. Calcium levels must be tightly regulated in a spatial and temporal manner in order to be efficiently and properly utilized in the host physiology. The Ca2+-ATPase
K Dukas et al.
Analytical biochemistry, 215(1), 66-72 (1993-11-15)
We developed a reverse transcription-polymerase chain reaction (RT-PCR) method which permits the simultaneous amplification of several mRNAs, even though their relative levels may be very different. First-strand cDNAs were synthesized from total RNA by MMLV reverse transcriptase with oligo(dT)15 priming.
Y M Zhu et al.
British journal of cancer, 91(11), 1970-1976 (2004-11-17)
Interleukin-8/CXCL8 (IL-8) is a chemokine and angiogenic factor. Recently, IL-8 was identified as an autocrine growth factor in several human cancers. Here, we investigated the expression and function of IL-8 in lung cancer cells. The expressions of IL-8 and its
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Articles

Reverse Transcription

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.

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RT-qPCR – Quantitative Reverse Transcription PCR

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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