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WTA2

Sigma-Aldrich

Complete Whole Transcriptome Amplification Kit

DNA polymerase included, Complete Kit with optimized enzyme to amplify total RNA in <4 hours, no 3′ bias

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Synonym(s):
transcriptome amplification kit
UNSPSC Code:
12352200
NACRES:
NA.55

Quality Level

200

technique(s)

whole genome amplification: suitable

shipped in

wet ice

storage temp.

−20°C

Related Categories

General description

WTA2 is optimized to amplify RNA from formalin-fixed, paraffin-embedded (FFPE) and other damaged or degraded samples. Whole Transcriptome Amplification (WTA) technology, allows for representative amplification of low nanogram quantities of total RNA in less than 4 hours without 3′-bias. Amplification products are suitable for applications such as qPCR, micro array analysis, and cloning. The WTA2 kit contains the polymerase needed to amplify the cDNA library.

Application

Complete Whole Transcriptome Amplification Kit is used for the following applications:
  • To establish a protocol for the simultaneous analysis of DNA and RNA viruses present in pig feces using process controlled deep sequencing.
  • Reverse transcription and cDNA amplification
  • For the synthesis and amplification of cDNA library using Genomic RNA released from immunocaptured PPV particles
  • Nucleic Acid Preparation and Deep Sequencing (The extracted nucleic acids were randomly primed for cDNA synthesis)
Suitable for use with downstream applications including:
  • qPCR
  • microarray analysis
  • cloning

Features and Benefits

  • Achieve up to 10,000x amplification in less than 4 hours with less than 30 minutes of "hands on" time required
  • Only 20 pg of total RNA template is required to amplify suitable cDNA for microarray profiling
  • Contains all needed components for cDNA amplification
  • Achieve linear amplification of expressed genes and exons without 3′ or 5′ bias
  • Effectively amplifies single cell or low input RNA, including mRNA and total RNA from any animal, plant, or microorganism

Principle

The WTA2 process involves two steps. In the first step, sample RNA is reverse transcribed with non-self-complementary primers composed of a quasi-random 3′ end and a universal 5′ end. During this process, displaced single strands serve as new templates for primer annealing and extension. The resultant cDNA library, comprised of random, overlapping 100 - 1000 base fragments flanked by universal end sequence. The 2nd step amplifies the cDNA library by PCR using WTA2 polymerase and a universal end primer to produce WTA2 product.

Kit Components Also Available Separately

Product No.
Description
SDS
  • Library Synthesis Enzyme
  • Library Synthesis Solution
  • Amplification Mix
  • Library Synthesis Buffer
  • W4502Water, Nuclease-Free Water, for Molecular BiologySDS
  • Amplification Enzyme
  • D7295Deoxynucleotide Mix, 10 mM, Molecular Biology ReagentSDS

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Regulatory Information

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Jana Sachsenröder et al.
PloS one, 9(2), e88888-e88888 (2014-03-04)
The pig faecal virome, which comprises the community of viruses present in pig faeces, is complex and consists of pig viruses, bacteriophages, transiently passaged plant viruses and other minor virus species. Only little is known about factors influencing its general
137 differential gene expression of in vitro-matured bovine oocytes with or without a polar body.
M. M. Pereira et al.
Reproduction, Fertility, and Development, 24(1) , 181-181 (2011)
Meik Dilcher et al.
Virology journal, 12, 183-183 (2015-11-07)
Rhabdoviridae infect a wide range of vertebrates, invertebrates and plants. Their transmission can occur via various arthropod vectors. In recent years, a number of novel rhabdoviruses have been identified from various animal species, but so far only few tick-transmitted rhabdoviruses
Aged stem cells reprogram their daily rhythmic functions to adapt to stress
Solanas G, et al.
Cell, 170, 678-692 (2017)
A Bal et al.
BMC infectious diseases, 18(1), 537-537 (2018-10-31)
In recent years, metagenomic Next-Generation Sequencing (mNGS) has increasingly been used for an accurate assumption-free virological diagnosis. However, the systematic workflow evaluation on clinical respiratory samples and implementation of quality controls (QCs) is still lacking. A total of 3 QCs
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