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The eBgDy™ Terminator v3.1 Cycle Sequencing Kit is used for performing Sanger sequencing on DNA samples. It is designed to provide high-quality sequencing data with high accuracy and minimal noise.
eBgDy™ Terminator v3.1 Cycle Sequencing Kit is a direct replacement for Applied Biosystem's BigDye™ Terminator v3.1 Cycle Sequencing Kit. Current BigDye™ v3.1 users can switch to eBgDy™ v3.1 seamlessly without changing current protocols. No new software or mobility shift file needs to be installed. No new calibration or validation is needed either.
The kit includes eBgDy™ Terminator v3.1 sequencing mix, which contains a proprietary mixture of dideoxynucleotide terminators labeled with different fluorescent dyes. The mix also includes DNA polymerase, which extends the primer annealed to the DNA template and incorporates the fluorescently-labeled dideoxynucleotide terminators, leading to chain termination and synthesis of DNA fragments of varying lengths.
The sequencing reaction is typically performed in a thermal cycler, which uses a series of temperature cycles to control the annealing and extension of the primers. After the reaction is complete, the products are purified and loaded onto a capillary electrophoresis instrument, such as the Applied Biosystems 3730xl DNA Analyzer, which separates the fragments by size and detects the fluorescent signals.
eBgDy™ Terminator v3.1 Cycle Sequencing Kit has the following features:
- High accuracy: The kit provides high-quality sequencing data with high accuracy and minimal noise, making it suitable for a wide range of applications, including de novo sequencing, variant detection, and genotyping.
- Wide range of applications: The kit can be used for sequencing various DNA templates, including PCR products, plasmids, BACs, and fosmids.
- Easy to use: The kit is designed to be easy to use, with a simple reaction setup and standardized protocols.
eBgDy™ v3.1 also brings many advantages over BigDye™ v3.1
- Faster reaction cycle: 3 min extension time, instead of 4 min, is need for eBgDy™, which can be further reduced to 2 min if needed.
- Higher dilution factor: As little as 0.15µl of eBgDy™ can be used with success in a 5 µl total volume reaction (Fig. 1)
- Higher success rate: Especially for difficult templates (Fig. 2)
Fig. 1 Top panel: Raw sequencing signal of a normal template using 0.15 µl BigDye™ v3.1 in duplicate. Bottom panel: Raw sequencing signal of the same normal template using 0.15 µl eBgDy™. Total reaction volume was 5 µl.
Fig. 2 Top panel: Raw sequencing signal of a GT rich template using 0.15 µl BigDye™ v3.1 in duplicate. Bottom panel: Raw sequencing signal of the same GT rich template using 0.15 µl eBgDy™. Total reaction volume was 5 µl.