GSM1971599: Sample_7P1641_WT_TA; Mus musculus; RNA-Seq
Muscles were removed, flash frozen and RNA was harvested using Trizol reagent. RNA libraries were prepared for sequencing using standard Illumina protocols A NGS library is made up of random fragments that represent the entire sample. It is created by shearing DNA (Covaris S220) into 150-400 bp fragments. These fragments are ligated to specific adapters. Library fragments of the appropriate size are then selected (size is application dependent) and isolated.  Following a sample cleanup step, the resultant library is quantified by qPCR and checked for quality using the Agilent Bioanalyzer. The SMF has automated library preparation for most applications using the Beckman SPRIworks system. Cluster Generation: Library fragments are bound to a flow cell by hybridizing the fragments to a lawn of oligonucleotides complementary to the adapter sequences. Bound fragments are clonally amplified by bridge amplification to create millions of individual dense clusters of clones.  Cluster generation occurs in a closed environment on the Illumina cBOT instrument.