WGS of Q. suber: genome survey using 170bp inserts
Paired-end libraries were constructed using the BGI 'DNA 170bp-800bp insert size Library Construction Protocol': 1.5µg DNA is fragmented by Covaris, then use the Gel-Electrophotometric to test the fragmented DNA. The fragmented DNA is combined with End Repair Mix, incubate at 20? for 30 min. Purify the end-repaired DNA with QIAquick PCR Purification Kit(Qiagen), then add A-Tailing Mix, incubate at 37? for 30 min. Combine the purified Adenylate 3'Ends DNA, Adapter and Ligation Mix, incubate the ligation reaction at 20? for 15 min. Adapter-ligated DNA is selected by running a 2% agarose gel to recover the target fragments. Purify the gel with QIAquick Gel Extraction kit (QIAGEN). Several rounds of PCR amplification with PCR Primer Cocktail and PCR Master Mix are performed to enrich the Adapter-ligated DNA fragments. Then the PCR products are selected by running another 2% agarose gel to recover the target fragments. Purify the gel with QIAquick Gel Extraction kit (QIAGEN).