PacBio Sequel Sequencing

Sequel System: high-throughput, cost-effective access to SMRT Sequencing

The Sequel System is based on Single Molecule, Real-Time (SMRT)

technology and is ideal for rapidly and cost-effectively generating high-quality

PacBiowhole genome de novo assemblies and full-length transcriptomes.

At GGBC, we provide all Sequel based services, from library prep and sequencing to data analysis.

Sequel System applications:

Consultation and Assistance

Contact for Genomics and Bioinformatics Consultation

Please use this link to send an email to Dr. Magdy Alabady for consultation on new or existing Genomics and Bioinformatics projects. Also, you can contact Dr. Alabady for assistance with grant proposals and for obtaining a letter of support from GGBC.

Contact for PacBio Technical Assistance

Please contact Roger Nilsen (rnilsen@uga.edu), Lab Manager, for technical questions about existing or new Sequel projects.

More contacts

Please visit our “who to contact?” page for detailed information about who you should contact at GGBC to receive a quick and accurate response.

Sample Preparation

High-quality, high-molecular-weight genomic DNA is crucial for obtaining long read lengths and optimal sequencing performance The SMART library preparation process does not utilize amplification techniques and resulting library molecules are directly used as templates for the sequencing process. The quality of the DNA and RNA starting material will be directly reflected in the extent of sequencing success or failure. Any unrepaired or irreversible DNA damage present in the input material (e.g., interstrand crosslinks, nicks, etc.) will result in impaired performance in the system.

Required quantitation method:

For DNA, please use pico green or equivalent fluorometric method (e.g., QUBIT).

For RNA, please use either ribo-green or QUBIT.

DNA quality: DNA should be high molecular weight (significantly higher than the desired insert size in the library) without any smear of degradation products.

RNA quality: Good quality, intact RNA is essential. RIN must ≥8.0 as indicated by the Agilent Bioanalyzer.

Buffer and concentration:  DNA or RNA should be in 10 mM Tris, pH 7.5-8.0 at a minimum concentration of 50-100ng/ul.

Required DNA and RNA quantities and concentration:

Sample concentration = 50-100 ng/ul as quantitated by QUBIT

Table 1. Sample Requirements

Library Size and Type Amount Required 
DNA10-20, 20-30 Kb
1-5 Kb

250-500 bp
DNA


Amplicons
7ug
4ug

500-750ng
RNA (IsoSeq)1-2, 2-3, 3-6, 6-10 Kb cuts

No cuts
Total RNA2ug

2ug


Prices and Quotes

Contact for Financial Inquiries and Quote Requests

Please email Dr. Myriam Bélanger, Operations Manager, for financial inquiries or to request a quote. Be as specific as possible, so that she can more quickly assist you.

Table 1. PacBio Sequel library preparation and SMRT cell run fees

ServiceUGA FeeNon-UGA FeeCommercial Fee
Large insert library (15 Kb-20 Kb)$670$791$1,005
Small fragments library (500 bp-4 Kb)$670$791$1,005
Library from amplicons (500 bp-4 Kb)Please inquirePlease inquirePlease inquire
Iso-Seq library$735$868$1,103
Barcoding for multiplexingPlease inquirePlease inquirePlease inquire
PacBio Sequel SMRT cell run (10 hour-movie per run)$1,150$1,357$1,725
Data Retrieval
  1. Primary analysis files will be transferred to customers through the GGBC’s data distribution FEX server. The following files are an example of the primary analysis output files: • *.adapters.fasta • *.scraps.bam.pbi • *.subreads.bam • *.subreadset.xml • *.txt • *.scraps.bam • *.sts.xml • *.subreads.bam.pbi • *.transferdone.

 

  1. The primary analysis files will be permanently deleted from GGBC’s data storage after 6 months from the data transfer date.