PCR-cDNA Barkoding Kit
PCR-cDNA Barkoding Kit
This kit is recommended for users who:
- wish to multiplex samples to reduce price per sample
- have a limited amount of input material
- want to optimise their sequencing experiment for throughput
- would like to identify and quantify full-length transcripts
- are interested in differential gene expression or differential transcript usage
- want to characterise and quantify isoforms, splice variants and fusion transcripts
Key features:
Prep time | 165 minutes |
Input amount | 1 ng poly-A+ RNA, or 50 ng total RNA |
Read length | Enriched for full-length cDNA during PCR |
Typical throughput | 7-12+ million full-length mapped reads in 48 hours per flow cell on MinION/GridION; 50+ million mapped reads in 48 hours per flow cell on PromethION |
Workflow
Taking full-length poly-A+ RNA, complementary strand synthesis and strand switching are performed using kit-supplied oligonucleotides. The kit contains 12 primer pairs which are used to generate and then amplify double-stranded cDNA by PCR amplification: each primer pair contains a barcode and 5’ tag which facilitates the ligase-free attachment of Rapid Sequencing Adapters. Amplified and barcoded samples are then pooled together, and Rapid Sequencing Adapters are added to the pooled mix.
