Unfiltered genomic sequences of SP70-1143- 200 bp libraries
For each library, 3-5 ug of genomic DNA was sheared using Covaris S220 Adaptive Focused Acoustics ultra sonicator. Libraries were constructed following standard protocol using the NEBNext DNA Sample Prep Master Mix Set 1 (NEB E6040) and Illumina-compatible paired-end adaptors. The unfiltered library was submitted to the same procedures of the digested library. Instead of McrBC endonuclease was added glicerol on the reaction. After 16 hr of incubation, undigested (Unfiltered –UF) library was purified using the QIAquick PCR Purification kit (Qiagen 28104), run on 2% MetaPhor*agarose (Lonza 50108) gel and fragments of desired sizes were excised from gel, purified with the QIAquick PCR Purification kit and amplified with 15 cycles of PCR using Phusion polymerase (part of the NEBNext kit) and cycling parameters described in the NEBNext kit manual. DNA concentrations were quantified on a Bioanalyzer (Agilent), diluted to 10 nM and loaded on flow cells to generate clusters.
Universidade Federal do Rio de Janeiro
200bp_unfiltered