Pacific Biosciences (PacBio) Third Generation Sequencing

PacBio Applications:

• De Novo Assembly: To finish genomes in order to fully identify, annotate and decipher genomic structures.
(www.pacb.com/denovo)

• Targeted Sequencing: To comprehensively characterize genetic variations including structural variants, haplotypes, and rare SNPs. (www.pacb.com/target)

• Base Modification Identification: Using real-time kinetic data to characterize genetic regulation and DNA damage. (www.pacb.com/basemod)

 

Library Prep Type	Sample Input Required	~ Average Insert	~ Mbp SMRTCell
10 kb MagBead*	1-10 ug	6,000 bp**	100-200 Mbp
10 kb CLR	5-10ug	5,000 bp	100-200 Mbp
5 kb CLR	2-4 ug	4,000 bp	100-200 Mbp
1kb CCS	500 ng - 1ug	500 bp	40 Mbp
500 bp CCS	250 ng - 1ug	350 bp	20 Mbp
250 bp CCS	250 ng - 1ug	150 bp	7.5 Mbp

 

*CLR: Continuous Long Read
*CCS: Circular Consensus Sequence (Note: CCS Mbp estimates are based on nonredundant coverage.)
**MagBead: PacBio MagBead sequencing provides more reads at longer lengths (comparative to a standard PacBio CLR library) and   reduces the sample input significantly.

Table Note: The standard result estimates outlined in the table above are based on output averages of several different projects and are to be used as a guideline. Individual project results may vary from these estimates. PacBio read-lengths and read quantities per SMRTCell are highly dependent on sample quality.

Quality Control: NCGR is committed to providing the best possible library prep and sequencing results for every sample we process. NCGR QC-tests all samples via Qubit (quantity) and Bioanalyzer and/or Gel (quality). 

PacBio Turnaround Time: Approximately four to eight weeks after receipt of signed quote, completed "NCGR Sample Form" documentation, and experimental samples of adequate sequencing quantity and quality.

XL Chemistry: NCGR offers PacBio sequencing using the new and improved XL polymerase chemistry and V3 SMRTCells resulting in longer read-lengths (averaging 5,000 bp) and an enhanced sequencing output of up to approximately 250 Mbp per SMRTCell.