Zymo D6331 PacBio Ultra-Low Input Library
Ultra-Low Input: DNA was extracted using the Shoreline Breaker Kit. A total of 10 ng of starting DNA was used to perform the PacBio "Preparing HiFi SMRTbell Libraries from Ultra-Low DNA Input" protocol (https://www.pacb.com/support/documentation/). DNA shearing was performed with a Megaruptor 3, sheared at speed 31. Typical steps were performed (remove single-strand overhangs, DNA damage repair, PCR adapter ligation), and the library was PCR amplified using 13 cycles for the two required reactions. Amplified DNA underwent end repair prior to SMRTbell adapter ligations. Size selection was performed using 3.7X Diluted AMPure. The resulting library was sequenced on a SMRT Cell 8M on the Sequel II System, with a 30 hr movie. The subreads were used to generate HiFi (CCS) reads using a minimum 99% accuracy cutoff. The gDNA amplification adapters were trimmed, and PCR duplicates were removed.
SUB8612752
Zymo_D6331_ULI