BigDye® Direct Cycle Sequencing Kit

Catalog number: 4458688

Applied Biosystems™  Related applications: Sanger Sequencing

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The BigDye® Direct Cycle Sequencing Kit, part of the BigDye® product family, is a complete kit for your PCR and Sequencing needs. This innovative, time-saving kit combines the PCR Purification and Cycle Sequencing Steps, enabling a streamlined protocol and shorter time to results. The kit is available in 24, 100, and 1000-reaction sizes to fit your sequencing throughput needs.

The BigDye® Direct Cycle Sequencing Kit Gives You These Advantages:
• A streamlined, simple procedure
• Performance of all sequencing steps in a single reaction tube
• Fast migration with excellent resolution starting at the 1st base after the gene-specific primer sequence
• Substantial time savings versus previous workflows

Save Time and Get Better Results with a Unique Streamlined Workflow
The BigDye® Direct Cycle Sequencing Kits have been developed to minimize complexity and hands-on time. Compared with previous sequencing procedures, which typically require 5 processing steps and take up to 8 hours, Big Dye® Direct can deliver results in 4 steps taking approximately 5 hours—up to a 40% time savings (Figure 1).

The kit provides reagents for 2 key steps of the sequencing workflow:
• The PCR reaction – using the BigDye® Direct PCR Master Mix and M13 tailed primer (not provided); PCR is performed in approximately 1 hour.
• The unique one-step PCR purification-cycle sequencing reaction – using the BigDye® Direct Sequencing Master Mix and the BigDye® Direct M13 primers provided (M13 tailed primer used for the PCR reaction simplifies the sequencing reaction setup by using 2 sequencing primers independently of the PCR product analyzed).

A One-Tube Protocol Gives You More Confidence
With the BigDye® Direct Cycle Sequencing Kit, all steps from PCR to loading on the capillary can be performed in the same reaction tube without transferring material from one tube to another. By minimizing the number of manual handling steps, the possibility of experimental variability is reduced; therefore, you can have greater confidence in your data.

An Integrated Solution Ideal for Resequencing
The BigDye® Direct Cycle Sequencing Kit is the first product designed to maintain conditions leading to high quality data generation throughout the entire read length with faster run times compared to common sequencing workflows. This is achieved through a unique reagent formulation that allows high-resolution data reads closer to the primer and the use of POP-7™ polymer for faster capillary electrophoresis times (Figure 2). The BigDye® Direct Cycle Sequencing Kit is optimized to work with Applied Biosystems® Genetic Analyzers and the BigDye XTerminator® Purification Kit.

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use
For Research Use Only. Not for use in diagnostic procedures.


For Use With (Equipment): GeneAmp™ 9700, Veriti® Thermal Cycler, 3500xL Genetic Analyzer, 3500 Genetic Analyzer, 3130xl Genetic Analyzer, 3130 Genetic Analyzer, 3730xl DNA Analyzer, 3730 DNA Analyzer
Recommended Applications: AT-Rich Sequencing (>65%), Comparative Sequencing (Germline Mutations - 50:50), GC-Rich Sequencing (>65%), GT-Rich or Difficult Template Sequencing, Gap Closure, Comparative Sequencing (Germline Mutations - 30:70 Heterozygotes), Standard Sequencing, Homopolymer A or T, Comparative Sequencing (Germline Mutations -10:90 Heterozygotes)
Format: 8-well strips, 96-well plate
Technique: Fluorescent Dye Terminator Sequencing
Product Size: 1,000 reactions
Template Compatibility: BAC DNA, Cosmids, Lambda DNA, PCR Amplicons (30:70 Heterozygotes), Fosmids, Plasmid DNA (≤15Kb), PCR Amplicons (50:50 Heterozygotes), Genomic DNA (Bacterial), PCR Amplicons (10:90 Heterozygotes)
Shipping Condition: Dry Ice

Contents & storage

This kit contains BigDye® Direct PCR Master Mix, Sequencing Master Mix, M13 Fwd Primer, M13 Rev Primer, and Control DNA CEPH 1347-02. Store all components at -15 to -25°C. PCR Master Mix should be stored at 4°C after first use. This kit contains sufficient reagents for 1000 reactions.


Manuals & protocols