ORDERING INFO |
Gene Knock Out ID
Plates with Custom Fimers
No Errors BAC sequencing with D-Strap Fimers NCI, NIH
(2004) PNAS coming soon
Closing the Gaps on Human Chromosomes (2004) Genome Research 14, 239-246
Direct BAC Sequencing with ThermoFidelase and Fimers
(2004) BACs 1, 295-308
(2004) BACs 1, 221-230
Production of Fimers for Whole Genome Direct Sequencing
(2004) Oligonucleotide Synthesis. Methods and Applications. 291-304
The B016 kit contains ThermoFidelase 2 and two Fimers for end sequencing from plasmid templates. It is designed for high throughput sequencing in 5 uL volume reactions with reduced consumption of dye terminator premix. Other versions of ThermoFidelase 2 kits with Fimers are designed for BAC, phage or genomic DNA sequencing.
ThermoFidelase 2 is a thermostable DNA unlinking enzyme (type IB DNA topoisomerase). During initial heating of the reactions it removes topological linkage between two strands of plasmid DNA. Because of the topological constraint in the DNA produced by unlinking, the unlinked plasmid strands cannot reform a normal double helix at the lower temperature used for primer annealing and extension. This results in an increased availability of targets for primer annealing and consequently a higher yield of cycle sequencing reaction products.
Fimers are primers with proprietary chemical modifications that inhibit unwanted reactions during cycle sequencing (e.g., primer-dimer artifacts or non-specific PCR). Specially designed Fimers have been successfully used for high throughput sequencing of difficult templates and highly sensitive reactions from minimal amount of templates (1 ng plasmid, 10 ng BAC, 100 ng microbial genomic DNA). Fimers can be designed for any sequencing applications. Kit B106 contains Forward and Reverse Fimers optimized for plasmids containing inserts of AT-rich Plasmodium falciparum DNA.
Template DNA, Dye Terminator Ready Reaction Mix, Deionized H2O
For sequencing of less than 96 samples, use the following table for calculation of the amount of each component
Covered by U.S. Patents 5,427,928, 5,656,463, 5,902,879, 6,548,251. Patents pending. Fidelase, ThermoFidelase, D-Strap and Fimer are trademarks of Fidelity Systems. ThermoFidelase, D-Strap and Fimer development is supported in part by the National Institutes of Health (NIGMS) and US Department of Energy