There are several options for bulk RNA-Seq and small RNA libraries:
RNA-Seq, Eukaryotic species:
mRNA-enriched: these libraries are constructed by first selecting polyA+ mRNAs, converting the mRNAs to cDNA, performing adaptor ligation and amplifying for the minimum number of PCR cycles required. At least 50ng of total RNA having a RIN > 7 and free from contaminating DNA should be submitted.
rRNA-depleted: for characterization of polyA+ and polyA- transcripts, libraries can be constructed by depleting rRNA and converting the leftover RNA to cDNA, instead of capturing polyA+ mRNAs.
FFPE samples: RNA-Seq libraries can be constructed from degraded and very low amounts of RNA by first converting all the RNA to cDNA and then using specific probes to remove rRNA, or by using probes that capture specific transcripts.
IsoSeq = full-length polyA mRNAs are sequenced in the PacBio Revio with high accuracy. This is the best way to fully characterize an entire transcriptome, including all splice-variants.
RNAseq, Microbial and Metagenomic samples:
rRNA-depletion: removal of rRNA can be done with probes that recognize microbial rRNAs or with probes that remove both host and microbial rRNAs. The leftover RNA is converted to RNA-Seq libraries that are individually barcoded with Unique Dual Indexes.
Small RNA, Eukaryotic species:
Libraries are constructed from 10ng of enriched small RNA fraction or 100ng of total RNA. RNAs from 15nt to 30nt in length are enriched by size selection on a PAGE gel.
Small RNA, bacterial:
Total RNA is treated with Antarctic Phosphatase and PNK, adaptors are added and RNAs 15nt to 250nt are enriched by size selection on a PAGE gel.
Circular RNAs, double-stranded RNAs:
We have extensive experience in the construction of these libraries.
