Biotechnology
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2000 Bases per Sample with Bi-Directional Sequencing
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Schematic of bi-directional sequencing on a double-stranded template. |
LI-COR's Simultaneous Bi-directional Sequencing (SBS™) on the 4300 System saves 50%, or more, or reagents and staff time. In bi-directional reactions, both forward and reverse primers are labeled with different infrared dyes and combined in one reaction.
SBS reactions produce over 1000 bases from each end of the template. Systems with 500 base read lengths can require four times as many templates to produce the same results as one SBS reaction.
Single Reaction Bi-directional Strategies
The Best Approach to cDNAUsing Standard labeled primers, both ends of cDNA clones can be sequenced simultaneously. Many average length cDNAs (approx. 2kb) can be completely sequenced in one SBS reaction, eliminating the time and expense of custom primers. Mutation ConfirmationBoth strands of most PCR products can be completely sequenced with SBS. Automatic analysis of opposite strand data with AlignIR can immediately confirm a heterozygote mutation sequence.
Images from the SBS reaction shown above identified a mutation in the "forward" reaction on the IRDye® 700 infrared dye image (left), and confirmed the mutation in "reverse" reaction from the IRDye® 800 infrared dye image (right). Similarly, bi-directional reactions can quickly resolve compressions and other difficult regions. |
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Genomic Clones
Using standard primers, both ends of BAC genomic clones can be sequenced in the same reaction. Long reads simplify sequence assembly.
