General descriptionWTA2 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.
ApplicationSuitable for use with downstream applications including:
- qPCR
- microarray analysis
- cloning
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)
- Amplification of RNA andssDNA viruses for Viral metagenomics from low-input clinical samples
Biochem/physiol ActionsThe 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.
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
