Ostrinia nubilalis lab population microbial midgut metagenome
Preparation of the DNA library for Shotgun sequencing consists of  fractionation of genomic DNA (gDNA) into smaller fragments (300-700 bp) in a  process called “nebulisation”. These fragments are subsequently polished  (blunted) and then, short Adaptors (A and B) are ligated onto both ends.  These adaptors provide priming sequences for the amplification and the  sequencing of the sample-library fragments. Adaptor B contains a 5'-biotin  tag that enables immobilization of the library onto streptavidin coated  beads. After nick repair, the non-biotinylated strand is released and used  as a single-stranded template DNA (sstDNA) library. The sstDNA library is  assessed for its quality and the optimal amount (DNA copies per bead) needed  for emPCR™ is determined by titration.