Products

DNA/RNA sequencing -PacBio Sequencer

PacBio sequencing platform is a long-read sequencing platform, which is also known as one of the Third-Generation Sequencing(TGS) technologies. The core technology, single-molecule real-time(SMRT), empowers the generation of reads with tens of kilo-base in length. On base of “Sequencing-by-Synthesis”, single nucleotide resolution is achieved by Zero-mode waveguide(ZMW), where only limited volume at the bottom(site of molecule synthesis), is illuminated. In addition, SMRT sequencing largely avoids sequence-specific bias in NGS system, in that most of PCR amplification steps are not required in library construction process.

Platform: Sequel II, Revio

Service Details

Demo Results

Features

Two sequencing modes on PacBio sequencer: Continuous Long Read (CLR) and Circular Consensus Read (CCS)

Sequencing Mode

Library Size

Theoretical Data Yield (Per Cell)

Single-base Accuracy

Applications

CLR

20Kb, 30Kb, etc.

80 Gb to 130 Gb

Approx. 85%

De novo, SV calling, etc.

CCS

15-20 Kb

14 to 40 Gb/Cell (Sequel II)

70 to 110 Gb/Cell (Revio)

Depends on the samples

Approx. 99%

De novo, SNP/Indel/SV calling, Iso-Seq,

Comparison of the performance and features of Revio and Sequel II

Terms	Sequel II system	Revio system	Increase
Higher density	8 million ZMWs	25 million ZMWs	3x
Independent stages	1	4	4x
Shorter run times	30 hours	24 hours	1.25x
30X HiFi human genomes / year	88	1,300	15x overall

Service Advantages

● Over 8 years experience on PacBio sequencing platform with thousands of closed projects with various species.

● Fully equipped with the latest PacBio Sequencing platforms, Revio to guarantee sufficient sequencing throughput.

● Faster turn-around time, higher data yield and more accurate data.

● Contributed in hundreds of high-impact PacBio-based publications.

Sample Requirements

Sample Type

Amount

Concentration （Qubit®）

Volume

Purity

Others

Genomic DNA

Depend on Data Requirement

≥50 ng/μl

≥15μl

OD260/280=1.7-2.2;
OD260/230=1.8-2.5；
Clear peak at 260 nm，no contaminations

Concentration needs to be measured by Qubit and Qubit/Nanopore = 0.8-2.5

Total RNA

≥1.2μg

≥120 ng/μl

≥15μl

OD260/280=1.7-2.5;
OD260/230=0.5-2.5；no contaminations

RIN value ≥7.5

5≥28S/18S≥1

Service Workflow

Sample preparation

Library construction

Sequencing

Data analysis

Project delivery

Previous: OEM manufacturer Mrna Sequencing Service - Long non-coding sequencing-Illumina – Biomarker

Next: Illumina pre-made libraries

1.In-house Data Yield

Data generated from 63 CCS cells (from 26 species)

DATA-PacBio-CCS-15 Kb	Average	Max	Min	Median
Yield - subreads (Gb)	421.12	544.27	221.38	426.58
Yiled - CCS(Gb)	25.93	38.59	10.86	25.43
Polymerase N50	145,651	175,430	118,118	144,689
Subreads N50	17,509	23,924	12,485	17,584
CCS N50	14,490	19,034	9,876	14,747
Average length-Polymerase	67,995	89,379	49,664	66,433
Average length-Subreads	15,866	21,036	11,657	16,012
Average length-CCS	14,489	19,074	8,575	14,655

Data generated from 16 CLR cells (from 76 species)

DATA-PacBio-CLR-30Kb	Average	Max	Min	Median
Yield - subreads (Gb)	142.20	291.40	50.55	142.49
Polymerase N50	39,456	121,191	15,389	35,231
Subreads N50	28,490	41,012	14,430	29,063
Average length-Polymerase	22,063	48,886	8,747	21,555
Average length-Subreads	17,720	27,225	8,293	17,779

2.Data QC – Demo Statistics on data yield

Sample	ccs Reads Num	Total ccs Bases (bp)	ccs Reads N50 (bp)	ccs Mean Length (bp)	ccs Longest Read (bp)	subreads Bases (bp)	ccs rate(%)
PB_BMKxxx	3,444,159	54,164,122,586	15,728	15,726	36,110	863,326,330,465	6.27