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mRNA-Seq 8-Sample Prep Kit
mRNA-Seq 8-Sample Prep Kit
| System: | Genome Analyzer, iScanSQ |
|---|---|
| Technology: | Illumina Sequencing |
| Assay: | mRNA-Seq |
| Applications: | Sequencing-Based Transcriptome Analysis |
| Contents: | Contains reagents for preparing cDNA libraries for sequencing |
The mRNA-Seq 8-Sample Prep Kit is used to build libraries for single-read and paired-end sequencing on the Genome Analyzer. Sample preparation is straightforward, using standard molecular biology techniques and requiring minimal hands-on time.
Starting with 1 μg total RNA, mRNA is isolated using polyA selection. The mRNA is then fragmented and randomly primed for reverse transcription followed by second-strand synthesis to create double-stranded cDNA fragments. Ends are repaired with a combination of fill-in reactions and exonuclease activity to produce blunt ends.
An 'A'- base is added to the blunt ends followed by ligation to Illumina Paired-End Sequencing adapters followed by PCR amplification. These adapters contain unique sequencing primer hybridization sites.
Additional sequences complementary to the oligonucleotides in the flow cell are added to the adapter sequences with tailed PCR primers. This is followed by gel-based size selection and purification to create libraries for cluster generation.
- mRNA template libraries are sequenced just as any DNA sample using Illumina Sequencing technology
- Use the mRNA-Seq 8-Sample Prep Kit with a Standard Cluster Generation Kit and 36-Cycle Illumina Sequencing Kit
Applications
Transcriptome Analysis
Explore the transcriptome like never before. Characterize all transcriptional activity, coding and non-coding, in any organism without a priori assumptions using the Genome Analyzer and achieve an unprecedented view of the transcriptome.
Illumina's unique combination of long and short reads, strand specificity, and millions of counts allows you to annotate coding SNPs, discover transcript isoforms, identify regulatory RNAs, characterize splice junctions, and determine the relative abundance of transcripts.
Generate transcript profiles in a single day with the greatest daily system output. Align RNA sequencing reads across splice junctions and map to reference genomes. Identify and quantify both rare and common transcripts, with six orders of magnitude of dynamic range. Reveal the hidden world of non-coding RNA architecture without prior information.
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