Library construction for RNA-sequencing is rather diverse because it changes depending on the type of RNA you want to sequence(mRNA, Total RNA, etc) and the quality of the RNA sample. GHTF offers several different type of library construction options depending on the type and condition of the RNA sample, so please look below to see which type of library construction is best for your samples.
Poly-A Enrichment Library Construction
This type of library construction isolates mRNA from the total RNA sample via poly-A enrichment. mRNA is extracted by hybridizing the poly A-tail on mRNA with the poly-T oligos on magnetic beads. This method is best for mRNA sequencing because it positively selects for mRNA, while the other types of RNA are removed. However this method requires high quality Total RNA, with a RIN score greater than 8.
rRNA Depletion Library Construction
This type of library construction removes only the ribosomal RNA from the total RNA sample. This method leaves every other type of RNA behind, so all types of RNA, with the exception of ribosomal, will be sequenced. This is useful when the target transcripts do not have a poly A tail or if you want to sequence non-coding RNA. Because this type of library preparation works by negatively removing rRNA, the input RNA quality does not have to be as high as the poly A enrichment method.
Micro RNA and Small RNA Library Construction
GHTF offers Micro RNA and Small RNA library construction from total RNA through double sided bead size selection. However users must take care in ensuring that their RNA isolation method retains micro and small RNA.
Please contact GHTF if you have any questions about RNA-sequencing.