The 454 GS Junior Sequencer brings the power of next generation sequencing technology directly to the research laboratory bench top. The same proven long-read sequencing, 400bp+, performance as the Genome Sequencer FLX System, scaled to suit the needs of individual labs. Quickly proceed from DNA to data to discovery with a simple sample preparation workflow, overnight sequencing and data processing, and a dedicated suite of data analysis software.
It has a 99% single read accuracy up to 4 homopolymers. After 4 it starts to drop off; however, the consensus accuracy is maintained at 99% for 12 plus homopolymers.
("JS Junior: Instrument and Workflow." 454 Sequencing. N.p., n.d. Web. 10 Nov 2011. <http://www.gsjunior.com/instrument-workflow.php)
Currently offered by the Vironomics Core to trained internal clients
● Set-up PicoTiterPlate and sequence on GS Junior
● Shotgun sequencing OR Amplicon sequencing
● Reagent Procurement
● Provide protocols
● Library/amplicon quality and quantification equipment
● QC and Maintenance of the GS Junior
● Provide data in .fna (sequence of contigs, FASTA format), .qual (corresponding Phred equivalent quality score), .sff (stored flowgram data)
● 2 years of data back-up
What you are responsible for…
- Meeting guidelines for sequencing sample submission
- Understanding that though the 454 has 400bp+ read lengths, the technology of the instrument does not do well with homopolymers.
- Attend training and perform Library preparation (Optional for amplicon sequencing)
- Attend training and perform Emulsion-based clonal amplification (emPCR amplification)
Go to our ordering page at https://infoporte.unc.edu/cores/buy.php?cid=69. UNC clients will need to sign-in. At the bottom of the page select the "Vironomics: Sequencing Bench Schedule" under the Timeshares section. Please click on the desired day and request activity and time on the pop-up. Once all calendar requests are complete, please Checkout along the left side and proceed to fill in the relevant fields. (Note: Use object code 3918 at the end of the page) Finally, place the order. The Vironomics Core Manager will contact you with approvals.
● We recommend going to www.my454.com , Roche sequencing web-site, and registering for an account. This will give you access to online protocols, research guides, technical bulletins, etc. Note: You will need to use your .edu email for this, also registration approval can take several weeks. Please contact the Vironomics Core if you need materials before your access is granted.
● Keep technical support phone number handy, they are available for questions, support, and troubleshooting. Roche Technical Support phone number: 1-800-262-4911
● For data analysis questions or suggestions please contact Dongmei Yang at 919-843-5292 or email@example.com (please CC firstname.lastname@example.org)
Contact the Vironomics core for questions, scheduling, and sample submission guidelines. Please see fee descriptions for cost.
Vironomics Core Manager: Marcia Sanders, 919-843-5292, email@example.com