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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

Quality Level

usage

sufficient for 100 reactions

feature

dNTPs included
hotstart

application(s)

RT-PCR: suitable

color

colorless

input

purified RNA

shipped in

wet ice

storage temp.

−20°C

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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.

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.

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

Los componentes del kit también están disponibles por separado

Referencia del producto
Descripción
SDS

  • O4387Random nonamers 100 μL

  • O4387Anchored oligo(dT)23 primers 100 μL

  • B017410x reaction buffers 10x AccuTaq buffer

  • P219210 mM dNTP mix 10 x PCR buffer

  • W1754Nuclease-free water 4 x 1.5

Storage Class Code

12 - Non Combustible Liquids

Flash Point F

Not applicable

Flash Point C

Not applicable

Certificado de Análisis

Certificado de origen

Anita Abu-Daya et al.
Developmental biology, 349(2), 204-212 (2010-10-28)
While limb regeneration has been extensively studied in amphibians, little is known about the initial events in limb formation in metamorphosing anurans. The small secreted integrin ligand nephronectin (npnt) is necessary for development of the metanephros in mouse. Although expressed...
17-estradiol attenuates LPS-induced interleukin-8 production by human peripheral blood monocytes through estrogen receptor-activation
Malisorn N, et al.
African Journal of Pharmacy and Pharmacology, 4(11), 806-810 (2010)
Chelsea T Smartt et al.
The American journal of tropical medicine and hygiene, 81(2), 258-263 (2009-07-29)
Alterations in gene expression in the midgut of female Culex pipiens quinquefasciatus exposed to blood meals containing 6.8 logs plaque-forming units/mL of West Nile virus (WNV) were studied by fluorescent differential display. Twenty-six different cDNAs exhibited reproducible differences after feeding...
Transgenic chickpea expressing a recombinant human $\alpha$ 1-proteinase inhibitor ($\alpha$ 1-PI) driven by a seed-specific promoters from the common bean Phaseolus vulgaris (L.)
Mishra S, et al.
Plant Cell, Tissue and Organ Culture, 115(1), 23-33 (2013)
Nicotiana tabacum osmotic stress-activated kinase is regulated by phosphorylation on Ser-154 and Ser-158 in the kinase activation loop
Burza AM, et al.
The Journal of Biological Chemistry, 281(45), 34299-34311 (2006)

Artículos

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.

Protocolos

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

Long and Accurate PCR Amplification of DNA with RedAccuTaq® (D4812)

Protocol for high fidelity amplification of long PCR fragments up to 22kb from complex DNA mixtures and up to 40kb from simple DNA mixtures. Red dye allows direct loading of reaction on a gel. REDAccuTaq LA

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