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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

NACRES:
NA.55

Quality Level

application(s)

whole genome amplification: suitable

shipped in

wet ice

storage temp.

−20°C

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 faeces. (Simultaneous identification of DNA and RNA viruses present in pig faeces 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.

Los componentes del kit también están disponibles por separado

Referencia del producto
Descripción
SDS

  • Library Synthesis Enzyme

  • Library Synthesis Solution

  • Amplification Mix

  • Library Synthesis Buffer

  • W4502Water, Nuclease-Free Water, for Molecular Biology

  • Amplification Enzyme

  • D7295Deoxynucleotide Mix, 10 mM, Molecular Biology Reagent

Storage Class Code

10 - Combustible liquids

Certificado de Análisis

Certificado de origen

Richard J Hall et al.
Journal of virological methods, 195, 194-204 (2013-09-17)
The discovery of new or divergent viruses using metagenomics and high-throughput sequencing has become more commonplace. The preparation of a sample is known to have an effect on the representation of virus sequences within the metagenomic dataset yet comparatively little...
Anna Sheveleva et al.
Virus genes, 47(2), 385-388 (2013-07-03)
The near-complete (99.7 %) genome sequence of a novel Russian Plum pox virus (PPV) isolate Pk, belonging to the strain Winona (W), has been determined by 454 pyrosequencing with the exception of the thirty-one 5'-terminal nucleotides. This region was amplified using...
Distinct degenerative phenotype of articular cartilage from knees with meniscus tear compared to knees with osteoarthritis
Rai MF, et al.
Osteoarthritis and Cartilage, 27, 945-955 (2019)
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...
Zahedan rhabdovirus, a novel virus detected in ticks from Iran
Dilcher M, et al
Virology Journal, 12, 183-183 (2015)

Artículos

Modification of the WTA2 Amplification Product for Next Generation Sequencing

Transplex Whole Transcriptome Amplification (WTA2) exponentially amplifies RNA producing a double-stranded cDNA library while precisely maintaining differential levels of individual transcripts in test and reference samples.

Whole Transcriptome Amplification of RNA from Low Cell-Number Samples

The efficacy of amplification of small quantities of total RNA with the Complete Whole Transcriptome Amplification Kit (WTA2) was examined in this study.

Histone Modification and Chromatin Remodeling | Epigenetics

Epigenetic modifications are thought to occur through two key interconnected processes—DNA methylation and the covalent modification of histones.

Protocolos

Integration of Sigma® TransPlex® WTA and the Complete WTA2 Kits with the NimbleGen™ Expression Microarray Workflow

Amplification products generated by the TransPlex® WTA and Complete WTA2 kits are suitable for microarray target for expression analyses, and can be readily integrated into existing NimbleGen workflows.

Integration of Sigma® TransPlex® WTA and the Complete WTA2 Kits with the Affymetrix Microarray Workflow

TransPlex WTA amplification product is suitable as microarray target for expression analysis on the Affymetrix platform, and can be incorporated into existing Affymetrix workflows

Integration of Sigma® TransPlex® WTA and the Complete WTA2 Kits with the Illumina Microarray Workflow

Amplification products generated by the TransPlex® WTA and Complete WTA2 kits are suitable for microarray target for expression analyses, and can be incorporated into existing Illumina workflows.

Complete Whole Transcriptome Amplification Kit Protocol (WTA2)

WTA2, a Whole Transcriptome Amplification (WTA) method, allows for representative amplification of nanogram quantities of total RNA in less than 4 hours without 3-bias

Contenido relacionado

Whole Transcriptome Amplification FAQs

Transplex Whole Transcriptome Amplification FAQs on topics including whole transcriptome steps, RNA source, including archival fixed tissue, library purification, quantitation of the product and downstream applications

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