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Stop/Gel Loading Buffer-LC

Stop/Gel Loading Buffer-LC

货号

SF81015

规格

1.5 ml

存储温度

-20℃

产品详情

Applications

  • Automated sequencing of all DNA templates, including those containing difficult-to-sequence regions that cannot be resolved with other sequencing kits (Figs. 1-4).

SequiTherm EXCEL II™ Sequencing Kits for automated sequencers use the SequiTherm EXCEL II Polymerase, which improves sequencing performance with increased signal intensities and the ability to sequence through GC-rich regions. The SequiTherm EXCEL II DNA Sequencing Kits incorporate the use of betaine,* which enables sequencing of a broad range of templates, including those containing regions that are very difficult to sequence. Protocols are provided for both cycle and high-temperature isothermal (noncycle) sequencing.

The kits have been optimized for use with the following DNA sequencers: LI-COR™ 4000 and 4200 series Sequencers, NEN™ Global IR2 DNA Sequencers, ALF™ DNA Sequencers, and ABI PRISM™ DNA Sequencers using dye primer technology. Using primers labeled with standard sequencing dyes, the SequiTherm EXCEL II kits may also be used with ABI PRISM 373 and 377 series sequencers. The kits are not compatible with ABI capillary sequencers (ABI 310, 3100, or 3700 series) or with BigDye™ technology.

Two different kits are available for use with LI-COR™ and NEN™ sequencing equipment. The first kit (Cat. No. SE9101LC) contains Termination Mixes that give optimal read lengths up to about 1,000 bases and is designed for use on 25- to 41-cm gels. The second kit (Cat. No. SE9202LC) provides enhanced signal intensities in the resolution range of 1,000-1,400 bases and is ideal for users interested in obtaining the longest possible reads in conjunction with 66-cm gels, version 4.x of the BaseImagIR™ software, and all versions of e-Seq™ software.

Benefits

  • Sequence templates containing hairpin structures (Fig. 3), regions of high GC or AT (Fig. 1) content, areas of interstrand reannealing, or nucleotide repeats (Fig. 4).
  • Reduced background signal results in clearer, less ambiguous sequence data (Fig. 2).
  • Kits compatible with LI-COR, NEN, and ALF DNA sequencers (both dye-primer and internal label) and with ABI PRISM automated sequencers (dye-primer).
  • Optional isothermal protocol to sequence templates that are recalcitrant to cycle sequencing. Termination Mixes contain 7-deaza-dGTP to reduce band compression during gel electrophoresis.

Figure 4. Sequencing through highly problematic repeat regions. Supercoiled plasmids containing GA-dinucleotide repeats (A), GGCtrinucleotide repeats (B), or GAAA-tetranucleotide repeats (C) were successfully sequenced with the SequiTherm EXCEL™ II DNA Sequencing Kit using the standard cycle sequencing protocol and dye-labeled primers.

*Covered by issued and/or pending patents.

 Figure 1. Sequencing of a difficult template. An AT rich template was sequenced using a standard cycle sequencing kit (A) and the SequiTherm EXCEL™ II Kit-LC (B). As shown, several strong stops were eliminated and sequence was easily read using the SequiTherm EXCEL II Kit-LC. Samples were loaded in the order G,A,T,C. Data were visualized on a LI-COR® Model 4000 DNA Sequencer.
Figure 2. The SequiTherm EXCEL™ II Kit-LC greatly reduces nonspecific background. A plasmid DNA template was sequenced with a standard cycle sequencing kit (A) and with the SequiTherm EXCEL II Kit-LC (B). Background signal is greatly reduced using the SequiTherm EXCEL II Kit-LC. Samples were loaded in the order G,A,T,C. Data were visualized on a LI-COR® Model 4000 DNA Sequencer.

A: SequiTherm EXCEL II™ DNA Sequencing Kit-LC.

Figure 3A

B: ABI BigDye™ Terminator Cycle Sequencing Kit.
Figure 3. SequiTherm EXCEL II™ DNA Sequencing Kits provide complete, clear sequencing data when other methods fail. A supercoiled plasmid containing a putative 24-bp hairpin loop was successfully sequenced with the SequiTherm EXCEL II DNA Sequencing Kit-LC using the standard cyclesequencing protocol and IRD800-labeled primer. Data were visualized on a NEN® IR2 Model 4000L DNA Sequencer (Panel A). This same region was not resolved using the ABI BigDye™ Terminator Cycle Sequencing Kit. Data were visualized on an ABI PRISM® 310 Genetic Analyzer (Panel B).

 

Catalog No. Size
SequiTherm EXCEL™ II DNA Sequencing Kit - LC for 25-41 cm gels
SE9101LC 100 Sequences
For LI-COR® or NEN®Global IR2 Sequencers.
SequiTherm EXCEL™ II DNA Sequencing Kit - LC for 66 cm gels
SE9202LC 100 Sequences
For LI-COR® or NEN®Global IR2 Sequencers.
Contents: SequiTherm EXCEL™ II DNA Polymerase, 3.5X SequiTherm EXCEL™ II Sequencing Buffer, SequiTherm EXCEL™ II-LC Termination Mixes (G, A, T, C), Control Template, Stop/Loading Buffer, Sterile Deionized Water.
SequiTherm EXCEL™ II DNA Polymerase
SED52100 100 Reactions
Includes SequiTherm EXCEL™ II Sequencing Buffer.
Stop/Gel Loading Buffer, Basic Fuchsin and Bromophenol Blue Dyes
SF81015 1.5 ml
For LI-COR®, NEN®. Sufficient for 100 sequences.
Basic Fuchsin Dye
SA85025 2.5 ml
For ALF™, ABI PRISM®, LI-COR®, NEN®. Sufficient for 100 sequences.

 

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