BigDye™ Terminator v1.1 Cycle Sequencing Kit
BigDye™ Terminator v1.1 Cycle Sequencing Kit
Applied Biosystems™

BigDye™ Terminator v1.1 Cycle Sequencing Kit

This BigDye™ Terminator v1.1 Cycle Sequencing Kit is designed for specialty applications that require optimal basecalling adjacent to the primerRead more
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Catalog NumberQuantity
433744924 Reactions
4337450100 Reactions
43374511000 Reactions
43374525000 Reactions
Catalog number 4337449
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Quantity:
24 Reactions

This BigDye™ Terminator v1.1 Cycle Sequencing Kit is designed for specialty applications that require optimal basecalling adjacent to the primer and for sequencing short PCR product templates with rapid electrophoresis run modules.

  • Improve the quality of your results for a wide range of sequencing applications
  • Sequence challenging templates and sequences more successfully
  • Get longer, higher-quality reads with more uniform peak heights and optimal signal balance
  • Enhance your productivity and reduce costs

Tackle Tough Sequences

With the BigDye Terminator v1.1 Cycle Sequencing Kit you get a robust, highly flexible chemistry for a wide range of applications, including de novo sequencing and resequencing.

Get More Uniform Peak Heights, Improved Accuracy

With BigDye Terminator v1.1 kit's superior chemistry, you generate data with uniform peak heights and optimized signal balance-which results in longer, higher-quality reads. You also get more accurate base assignments for heterozygote and mutation detection.

Easily Integrate into Your Workflow

BigDye™ Terminator v.1.1 kit's chemistry requires no new software or instrument recalibration.You can easily integrate either kit into your current workflow with minimal changes to your protocols.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
For Use With (Equipment)SeqStudio™ Genetic Analyzer, SeqStudio™ Flex Series Genetic Analyzer, 3500/3500xL Genetic Analyzer, 3730/3730xl DNA Analyzer Thermal Cyclers: GeneAmp™ PCR System 9700 Dual Series, Veriti™ Fast 96‐Well and 384‐Well Thermal Cyclers
For Use With (Application)AT-Rich Sequencing (>65%), Comparative Sequencing (Germline Mutations - 50:50), Short PCR Product Sequencing, GC-Rich Sequencing (>65%), GT-Rich or Difficult Template Sequencing, De Novo Sequencing - Low to Mid Throughput
Product LineBigDye™ Terminator
Product TypeSequencing Buffer
Quantity24 Reactions
TechniqueFluorescent Dye Terminator Sequencing
Template CompatibilityBAC DNA, Plasmid DNA (≤15 Kb), PCR Amplicons, Single Stranded DNA
Unit SizeEach
Contents & Storage
• 200 μL Tube of BigDye™ Terminator v1.1 Ready Reaction Mix
• Tube M13 (-21) Primer
• Tube pREF-BDT™ Control DNA (200 ng/μL)
• 1 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.

Frequently asked questions (FAQs)

My DNA sequencing data from the capillary electrophoresis system does not look good. Why?

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.

•pREF-BDT Control DNA: validate cycle sequencing and its clean-up
If the pREF-BDT control reactions fail and the Sequencing Standards pass, look into potential problems with the sequencing kit, thermal cycler, and cleanup procedure.
The pREF-BDT 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.

What are the concentrations of the pREF-BDT and M13 primer controls that come with the BigDye Terminator kits?

The concentration of the pREF-BDT control is 200 ng/µL. The concentration of the M13 primer is 0.8 pmol/µL.

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

How do I determine the annealing temperature for the DNA sequencing thermal cycling conditions?



Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

Can I purchase the -21 M13 forward control primer from the BigDye Terminator kit separately?

We do not sell the -21 M13 forward control primer separately, however we provide the primer sequence for your convenience: TGTAAAACGACGGCCAGT

Please feel free to use our oligo services to order this primer (https://www.thermofisher.com/us/en/home/products-and-services/product-types/primers-oligos-nucleotides/invitrogen-custom-dna-oligos.html).
We also offer an -21M13 forward primer that is one basepair shorter than that contained in the kit (Cat. No. N52002).

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

Can I purchase the pREF-BDT control from the BigDye Terminator kit separately?

We do not sell the pREF-BDT control separately from the DNA sequencing kit.

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

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