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WTA2

Sigma-Aldrich

TransPlex® Complete Whole Transcriptom Amplification Kit

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

别名:
完整全转录组扩增试剂盒
NACRES:
NA.55

质量水平

200

应用

whole genome amplification: suitable

运输

wet ice

储存温度

−20°C

一般描述

经过优化的WTA2,可对福尔马林固定的石蜡包埋样品(FFPE)和其他受损或降解样本中的RNA进行扩增。全转录组扩增(WTA)技术可在4小时内对ng级的总RNA进行无3′偏差的代表性扩增。扩增产物适用于qPCR、微阵列分析和克隆等应用。WTA2试剂盒含有cDNA文库扩增所需的聚合酶。

应用

完整全转录组扩增试剂盒用于:
  • 建立同时分析猪粪DNA和RNA病毒的实验方案。(通过可控工艺深度测序同时鉴定猪粪中存在的DNA和RNA病毒)
  • 逆转录和cDNA扩增
  • 使用免疫捕获PPV颗粒中释放的RNA进行cDNA文库合成和扩增
  • 核酸制备和深度测序(提取的核酸使用随机引物进行cDNA合成)
适于各种下游应用,包括:
  • qPCR
  • 微阵列分析
  • 克隆

特点和优势

  • 在4小时内实现高达10,000x的扩增,并且手动操作时间低于30分钟
  • 仅需20 pg的总RNA模板,即可扩增出适用于微阵列分析的cDNA
  • 含有cDNA扩增所需的所有元件
  • 实现对表达基因和外显子的线性扩增,无3′或5′偏差
  • 有效扩增单细胞或小量RNA,包括来自动物、植物或微生物的mRNA和总RNA。

原理

WTA2过程包括2个步骤。第一步,使用由拟随机3′端和通用5′端组成的非自身互补引物对样品RNA进行逆转录。在该过程中,被取代的单链将作为引物退火和延伸的新模板。所得OmniPlex cDNA文库由以通用末端序列为侧翼序列、含100-1000个碱基的随机重叠片段组成。第二步,使用WTA2聚合酶和通用末端引物对cDNA文库进行PCR扩增,生成WTA2产物。

试剂盒组分也可单独购买

产品编号
说明
化学品安全说明书

  • 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

储存分类代码

10 - Combustible liquids

WGK

WGK 3

分析证书

原产地证书 (CofO)

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
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)
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)

技术文章

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.

实验方案

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

相关内容

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.

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

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.

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