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Home
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Biochemical Reagents
DNA Reagents
DNA Sequencing
BigDye™ Terminator v3.1 Cycle Sequencing Kit


Applied Biosystems™


BIGDYE™ TERMINATOR V3.1 CYCLE SEQUENCING KIT


Catalog number: 4337458
Related applications: DNA Sequencing

Sanger Sequencing

SARS-CoV-2 Pathogen Research Solutions
Technical SupportCustomer Service
Applied Biosystems™


BIGDYE™ TERMINATOR V3.1 CYCLE SEQUENCING KIT


Catalog number: 4337458
Related applications: DNA Sequencing

Sanger Sequencing

SARS-CoV-2 Pathogen Research Solutions
Technical SupportCustomer Service


Catalog Number
4337458
Unit Size
25,000 reactions
Price (CAD)
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Catalog Number
4337454
Unit Size
24 reactions
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Catalog Number
4337455
Unit Size
100 reactions
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Catalog Number
4337457
Unit Size
5,000 reactions
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Catalog Number
4337456
Unit Size
1,000 reactions
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Catalog NumberUnit SizePrice (CAD)

433745825,000 reactionsContact Us ›

433745424 reactionsContact Us ›

4337455100 reactionsContact Us ›

43374575,000 reactionsContact Us ›

43374561,000 reactionsContact Us ›

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Product Overview
Figures
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Recommendations
Recommendations
Documents
FAQ
Citations & References
Additional Information
Recommendations

The BigDye™ Terminator v3.1 Cycle Sequencing Kit's robust, highly flexible
chemistry is ideal for de novo sequencing, resequencing, and finishing with PCR
product, plasmid, fosmid, and BAC templates.

• Improve the quality of your results for a wide range of sequencing
applications
• Optimized for long read lengths
• Better dye mobility characteristics
• Improved performance reading through GT-rich regions
• Get longer, higher-quality reads with more uniform peak heights and optimal
signal balance
• Enhance your productivity and reduce costs

Get more uniform peak heights and improved accuracy
With BigDye™ Terminator v3.1 kit superior chemistry, you generate data with
uniform peak heights and optimized signal balance. This results in longer,
higher-quality reads and more accurate base assignments for heterozygote and
mutation detection.
For Research Use Only. Not for use in diagnostic procedures.


SPECIFICATIONS

Description
Cycle Sequencing Kit, 25,000 rxns, Includes 10 x 20mL tubes of BigDye™
Terminator v3.1 Ready Reaction Mix, 4 tubes M13 (-21) Primer, 4 tubes pGEM
Control DNA, 1 x 233mL tube of 5X Sequencing Buffer (may be stored at 4°C)
Format
96- or 384-well Plate, 8-well Strips, 12-well Strips
For Use With (Equipment)
310 Genetic Analyzer, 3130xl Genetic Analyzer, 3730xl DNA Analyzer, GeneAmp
9700, Veriti Thermal Cycler, 3130 Genetic Analyzer, 3730 DNA Analyzer, 3500xL
Genetic Analyzer, 3500 Genetic Analyzer
For Use With (Application)
Standard Sequencing
Includes
10 x 20mL tubes of BigDye™ Terminator v3.1 Ready Reaction Mix, 4 tubes M13 (-21)
Primer, 4 tubes pGEM Control DNA, 1 x 233mL tube of 5X Sequencing Buffer (may be
stored at 4°C)
No. of Reactions
25,000
Product Line
BigDye™ Terminator, BigDye™
Product Type
Sequencing Buffer
Technique
Fluorescent Dye Terminator Sequencing
Template Compatibility
BAC DNA, Plasmid DNA (≤15Kb), PCR Amplicons, Single Stranded DNA


CONTENTS & STORAGE

• 10 × 20-ml tubes of BigDye™ Terminator v3.1 Ready Reaction Mix
• 4 tubes M13 (-21) Primer
• 4 tubes pGEM Control DNA
• 1 × 233-ml tube of 5X Sequencing Buffer (may be stored at 4°C)

Store kit at -15°C to -25°C, and avoid repeated freeze-thaw cycles.



FIGURES






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 * 


DOCUMENTS & DOWNLOADS


CERTIFICATES

Search by lot number or partial lot number

Search


Request a Certificate


SAFETY DATA SHEETS

SDS



SCIENTIFIC RESOURCES

 * APPLICATION NOTES
   
   
   Application Notes & Tutorials: Faster Cycle Sequencing Applications Using the
   Applied Biosystems 9800 Fast Thermal Cycler: Application Note (English )
   

 * REFERENCE MATERIALS
   
   
   White Paper: Genetic analysis tools for genome editing workflows
   White Paper: Genetic analysis tools for genome editing workflows
   

 * POSTERS
   
   
   Poster: Direct Sequencing From Culture
   

 * BROCHURES
   
   
   Application Notes & Tutorials: A Workflow for Obtaining High Quality
   Sequencing Data from Bacterial Artificial Chromosome (BAC) DNA (English )
   Brochure: Sanger sequencing—Generate high-quality data with our proven
   workflow
   Amplifi cation of Long GC-Rich Sequences
   Product Bulletin: BigDye Terminators v3.1 and v1.1
   Application Notes & Tutorials: Reliable Resequencing of Human Mitochondrial
   Genome to Identify Mutations Linked to Mitochondrial Related Diseases
   (English )
   

 * VECTOR INFORMATION
   
   
   Additional Document: Longer, higher quality reads with BigDye® Terminator
   v3.1 and v1.1 kits
   


PRODUCT INFORMATION

 * MANUALS
   
   
   Product Sheet: Sequencing Standards, BigDye Terminator v3.1 - SeqStudio Flex,
   SeqStudio, and 3500 series instruments
   Product Sheet: BigDye Terminator v3.1 Matrix Standards Kit
   BDT3.1_CycleSeq-QRC(Chinese)
   Quick Reference: BigDye Terminator v3.1 Cycle Sequencing Kit
   User Bulletin: Using ExoSAP-IT Express PCR Product Cleanup to generate high
   quality BigDye Terminator v3.1 Cycle Sequencing Kit Data
   Fast Cycle Sequencing Using the 9800 Fast Thermal Cycler
   User Guide: BigDye Terminator v3.1 Cycle Sequencing Kit
   Quick Ref: BigDye® Terminator v3.1/v1.1 (Japanese)
   User Bulletin: Generating high-quality data using the BigDye Terminator v3.1
   Cycle Sequencing Kit Using CleanSweep PCR Purification
   User Bulletin: Generating high?quality data using the BigDye Terminator v3.1
   Cycle Sequencing Kit Using ExoSAP-IT Reagent for PCR purification
   





FREQUENTLY ASKED QUESTIONS (FAQS)

 * My DNA sequencing data from the capillary electrophoresis system does not
   look good. Why?
   Answer
   
   Capillary electrophoresis systems are complex systems, and troubleshooting
   data quality can be complicated. The biggest contributors to data quality can
   be divided into these areas: software, chemistry, and instrument performance.
   
   Troubleshooting software
   Ensure that the run module you are using and the dye set match the chemistry
   and instrument setup. For example, if you are running a multicapillary
   instrument with a 50 cm array on it, the run module should have a “50” in the
   name. On a multicapillary system, BigDye Terminator v3.1 should run under Dye
   Set Z, BigDye Terminator v1.1 chemistry should run in Dye Set E (on the ABI
   PRISM 310 Genetic Analyzer, both chemistries run under Filter Set E, but
   require different matrices for analysis). The mobility file (DyeSet/Primer)
   should have the correct instrument, polymer, and chemistry in the name (e.g.
   KB_3130_POP7_BDTv.3.mob).
   
   Troubleshooting chemistry and instrument performance
   •Sequencing Standards: validate instrument
   If Sequencing Standards fail, it suggests a possible electrophoresis problem
   due to array, polymer, buffer, water, septa, plastics, or maybe a hardware
   failure such as autosampler, laser, camera, etc.
   
   •pGEM Control DNA: validate cycle sequencing and its clean-up
   If the pGEM control reactions fail and the Sequencing Standards pass, look
   into potential problems with the sequencing kit, thermal cycler, and cleanup
   procedure.
   The pGEM Control DNA and M13 (–21) primers needed to run the reactions are
   included with each BigDye Terminator Cycle Sequencing Kit
   
   •Custom Internal Controls: validate PCR and its clean-up
   If the Sequencing Standards and the pGEM controls passed, then look into
   potential problems with template quantity and quality, primers, PCR reaction
   and purification.
   
   For more information on troubleshooting your data, please refer to the DNA
   Sequencing by Capillary Electrophoresis: Applied Biosystems Chemistry Guide:
   3rd Edition, which can be downloaded from this link
   (https://www.thermofisher.com/us/en/home/global/forms/sanger-sequencing-guide-download-form.html).
   
   Find additional tips, troubleshooting help, and resources within our
   Capillary Electrophoresis Applications Support Center.

 * I see another sequence running under mine. What can cause this?
   Answer
   
   Some of the causes of another sequence appearing under the primary sequence
   are:
   
   • Contamination: There is more than one species of DNA present (e.g.,
   multiple PCR products).
   
   • Primers: Primer is annealing in more than one location on the template,
   primer dimer, primer degradation or not manufactured properly resulting in
   N+1 or N-1, carry over from PCR reaction, or primers pairs in a multiplex
   reaction that are not appropriate for multiplexing (i.e., primers anneal
   inappropriately).
   
   • Spectral/Matrix: If the raw data signal intensity of the sample is too high
   or saturated it can exceed the amount of color bleedthrough (or spectral
   overlap) that the matrix (310) or spectral (3130, 3730, 3500) are removing,
   resulting in secondary peaks appearing in a very specific pattern (e.g., a
   red peak always appearing under a green peak). A change in the camera, laser,
   or optical alignment requires that a new matrix be made or a new spectral
   calibration be performed.
   
   For more information on more than one sequence or set of peaks in a
   sequencing run, please refer to the DNA Sequencing by Capillary
   Electrophoresis: Applied Biosystems Chemistry Guide: Second Edition (Cat. No.
   4305080, Rev. C). The guide can be found by searching the Thermo Fisher
   Scientific website with the catalog number 4305080.
   
   Find additional tips, troubleshooting help, and resources within our
   Capillary Electrophoresis Applications Support Center.

 * I ran out of 5X Sequencing Buffer. What is the formulation so I can make my
   own?
   Answer
   
   The formulation of the 5X Sequencing Buffer that comes with the BigDye
   Terminator Kit is proprietary.
   
   Find additional tips, troubleshooting help, and resources within our
   Capillary Electrophoresis Applications Support Center.

 * What information should I include when submitting data to Genetic Analysis
   Technical Support?
   Answer
   
   When contacting Genetic Analysis Support (FAS, FSE, Technical Support),
   please have (if possible):
   
   Instrument model and serial numbers
   Version of the software are you using
   Application and/or kit information
   Kit lot number
   Number of runs on the capillary or array
   Polymer type and lot number
   Buffer lot number
   Hi-Di Formamide lot number or loading solution information
   Sample data that can be sent to the support person
   Details on:
     -Reaction setup (i.e., how much DNA was used, primer, etc.)
     -How the reactions were cleaned up (alcohol precipitation, columns, etc)
     -What template DNA was used (i.e. plasmid, PCR product)
     -How the template DNA was prepared?
   
   All of this information will help the support person quickly and accurately
   assess the problem and provide you with recommendations for resolution.
   
   Find additional tips, troubleshooting help, and resources within our
   Capillary Electrophoresis Applications Support Center.

 * What is the difference between the regular BigDye (Terminator) kits and the
   BigDye Direct Cycle Sequencing Kit?
   Answer
   
   BigDye Direct Cycle Sequencing Kit supplies reagents for the complete cycle
   sequencing workflow, including the PCR reaction, reaction cleanup, and
   sequencing the PCR product. The post-PCR and cycle sequencing process is
   combined into a single step, and the workflow requires that the initial PCR
   primers be designed with an M13 tail, since M13 primers will be used for the
   sequencing reaction. BigDye Terminator Cycle Sequencing Kits are designed so
   that the initial PCR and the cleanup are performed as separate reactions.
   This allows the user more flexibility in primer design and the option to
   break up the workflow. For the comparative studies that we performed
   in-house, we found sequencing data obtained using these two kits to be
   concordant and of at least equivalent quality (typically the BigDye Direct
   Cycle Sequencing Kit produced better results).
   
   Find additional tips, troubleshooting help, and resources within our
   Capillary Electrophoresis Applications Support Center.

View more


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