MYreads Custom DNA and RNA Sequencing Services
PRICING: Due to the wide range of services offered and the possibility to combine several services into a single package, please contact us for a quote.
454 Sequencing Library Preparation Services
Genomic DNA Rapid Library Preparation
Genomic DNA sample are randomly fragmented to an average size of 600 to 900 bp following Roche 454 recommendations, and size-selected to discard short fragments. DNA fragments are end-repaired and ligated to 454 Rapid Library adaptors. At this step, any MID tag can be inserted to permit subsequent library pooling/multiplexing during sequencing. The library is quantified by fluorometry and the DNA fragment length distribution assessed on an Agilent Bioanalyzer. [See an example]
PCR Amplicon Library Preparation
Templates in the samples are PCR amplified using loci-specific primers introducing the sequencing adaptors at the same time. The PCR is performed in an emulsion to prevent over-amplification of particular sequences and sequence recombinations. Preferentially, the amplicon size(s) will be in the 200 to 600 bp. The amplicon library is quantified by fluorometry and the DNA fragment length distribution assessed on an Agilent Bioanalyzer. [See an example]
Sequence capture Services - Any Sequencing Platform
Sequence Enrichment by Capture
The sequencing library (454, Illumina, Solid, Ion Torrent) is denatured and hybridized to a collection of biotinylated RNA baits (see MYselect kits) complementary to the sequences being targeted. Targeted sequences hybridized to the baits are pulled out of the solution using magnetic beads. After several stringent washes, captured genomic fragments are released from the baits.
Sequence Enrichment by Depletion
The principle of sequence enrichment by depletion is that unwanted sequences are removed from the solution by hybridization to complementary baits. In this case, it is necessary to have baits targeting both strands for complete depletion. The sequencing library (454, Illumina, Solid, Ion Torrent) is denatured and hybridized to a collection of biotinylated RNA baits (see MYselect kits) complementary to both strands of the sequences to be depleted. Sequences hybridized to the baits are pulled out of the solution using magnetic beads, leaving your enriched sample in the solution.