Physical methods for large fragment sizing and sequencing using nanopore technologies and short fragments by fragmentation sequencing using mass spectrometry are also under rapid development of throughput, accuracy, and sensitivity.
The purified mRNA was then disrupted into short fragments using fragmentation buffer.
Briefly, eukaryotic mRNA was extracted from total RNA using oligo (dT) magnetic beads and cleaved into short fragments using fragmentation buffer.
We then enriched the poly (A) mRNA using beads with Oligo (dT) and interrupted mRNA into short fragments with fragmentation buffer.
The mRNA was sheared into short fragments with fragmentation buffer and selected for construction of Illumina RNA-Seq libraries according to the described protocol.
After the extraction of total RNA, oligo (dT) magnetic beads were used to isolate poly (A) mRNA, which can break mRNA into short fragments when fragmentation buffer is added.
The shorter fragmentation time was used to yield slightly larger library fragments of 350-400 bp.
All mRNA was broken into short fragments by adding fragmentation buffer.
The mRNAs were isolated by using oligo (dT) magnetic beads and was interrupted into short fragments by adding fragmentation buffer.
After these, magnetic beads with Oligo (dT) were used to isolate mRNA which was fragmented into short fragments using a fragmentation buffer.
