Sequencing Analysis Software v5.4 Initial License
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Applied Biosystems™

Sequencing Analysis Software v5.4 Initial License

This software enables you to basecall, trim, display, edit, and print data from our entire line of capillary DNA sequencingRead more
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Catalog NumberQuantity
43609671 Ea.
Catalog number 4360967
Price (CNY)
-
Quantity:
1 Ea.
This software enables you to basecall, trim, display, edit, and print data from our entire line of capillary DNA sequencing instruments for data analysis and quality control.

• Obtain longer read lengths, more high-quality bases, and increased accuracy at the 5' end
• Get increased accuracy in regions with low signal-to-noise ratios or with anomalous signal artifacts such as spikes or dye blobs
• Determine the quality of your data using superior metrics from basecalling quality values
• Accelerate quality control using analysis reports with analysis statistics
• Filter out low-quality sequence ends automatically with sequence trimming

Longer Read Lengths with High-Quality Base Pairs
Our significantly improved basecalling algorithm, the KB™ basecaller now gives you up to 100 more high-quality bases than other basecalling algorithms. You also get longer read lengths with high-quality base pairs, mixed basecalling with quality value, and accurate basecalling of usually difficult-to-sequence short PCR fragments.

Easily Review Sequencing Results with Quality Values
This software enables you to customize and color code the range of the quality values to represent low-, medium-, and high-quality bases. This way, when the basecaller identifies each base and assigns it a quality value, all you have to do is look at the color coding to easily review, discard, or accept it. In addition, the software trims the ends of low-quality bases, grays them out on the user interface for easy identification, and calculates a sample score, which is the average quality value for all the bases in the untrimmed region.

Reduce Data Screening Time
Eliminate manual review of sequencing data batches. With the software's Quality Control (QC) reports, you get read length and sample score (average QV of bases in the clear range) for each sample file, enabling you to sort data by quality. And to make reviewing data even easier, each QC report is hyperlinked back to its source data

For Research Use Only. Not for use in diagnostics procedures.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Data EntrySample files generated by all 310, 377, 3100, 3700, and 3730 Genetic Analyzers
Data OutputsABI (.ab1), FASTA (.seq), Phred (.phd.1), chromatogram (.scf), and Analysis Reports
For Use With (Application)Sequencing
LicenseStatic
Operating SystemWindows Vista Business, Windows XP Pro
Product TypeSequencing Analysis Software
Quantity1 Ea.
SoftwareSequencing Data Analysis
System RequirementsComputer: Core 2 Duo E8400/3 GHz; 2 GB RAM
Unit Size1 software
Contents & Storage
Contains one software CD.

Frequently asked questions (FAQs)

How do I make a sequencing matrix on the 310 Genetic Analyzer?

You can make a matrix from running the Sequencing Standard or by running Matrix Standards. To make the Matrix:

(1) After the run, open the DNA Sequencing Analysis Software.

(2) Go to the File menu and select Add Sample(s). Select the file(s) of the standards that you ran, click on the “Add Selected Samples” button, then click the OK box.

(3) After the samples have been added to the Sample Manager, click the Show box, and check the raw data. Peak height should be greater than 200 rfu and the baseline needs to be stable.

(4) Go to the “Tools” menu and select “Make Matrix”. Select the number of files you will be generating the matrix from (e.g., if you are generating it from a sample file, select 1; if from Matrix Standards, select 4).

(5) Click on the “…” box to the right of each line to browse to the sample file(s). Select the file and click “Open”. Do this for all of the samples you will use for the Matrix.

(6) In the “Specify the path for the new matrix file” area, name your matrix. The location the file should be stored in should be the default location and in D:\AppliedBio\Shared\Analysis\Basecaller\Matrix. Note:The matrix file needs to be in both locations in order to autoanalyze the data. If you attempt to point the preferences to look for the matrix in D:\AppliedBiosystems\SeqA5.X\AppSeqA\bin\Basecaller\Matrix, the entire pathway will be written to the .ab1 file for the matrix name and autoanalysis will fail.

(7) Click the “Make Matrix” button. The matrix will either be made correctly or it will generate an error message with specific information on why it failed.

(8) Either copy and paste the Matrix file from the default location to D:\AppliedBio\Shared\Analysis\Basecaller\Matrix or make the matrix 2, the second being saved to that location.

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

What do I need to do to get the Sequencing Analysis v.5.X Software to autoanalyze my 310 Genetic Analyzer sequencing samples?

In order to autoanalyze samples on the 310 Genetic Analyzer, there are three things that need to be done:

(1) Copy the Matrix and Mobility files from the Matrix and Mobility folders located at D:\AppliedBiosystems\SeqA5.X\AppSeqA\bin\Basecaller and paste them into the Matrix and Mobility folders at D:\AppliedBio\Shared\Analysis\Basecaller. If you have not made a matrix yet, you will need to do so prior to running samples.
(2) Open Sequencing Analysis, create an Analysis Protocol, and set it as the Analysis Default.
(3) Set up the Preferences to point to the mobility and matrix files in the local directory and in the General Settings tab, point the Autoanalyze with menu to D:\AppliedBiosystems\SeqA5.4\AppSeqA\Automation310.exe

Once the files are copied and the preferences set, you will be ready to run. At the end of the run, the sample files (.ab1) will be analyzed, but you will not see Sequencing Analysis open. The sample files are analyzed with a non-visible version of Sequencing Analysis.

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

What is the relationship between QV20 and the KB Basecaller and how do they determine the length of read specifications?

The improved basecalling algorithm (the KB Basecaller) is designed to accurately call more bases out of sequencing data, and reduce manual data review time by providing per-base quality value (QV) assignment, which is a measure of basecaller accuracy. The KB Basecaller supports most chemistries and run modules available on the Applied Biosystems 3130 Series Genetic Analyzers running Data Collection v3.0 and v.3.1 Software, and is also integrated in the Applied Biosystems Sequencing Analysis v. 5.X Software and SeqScape v2.X Software. In previous versions of Data Collection Software, resolution performance for all run modules was calibrated with a 98.5% basecalling accuracy (1.5% probability of error) specification using the ABI Basecaller. Utilizing the KB Basecaller v1.2 release for Data Collection v3.0 Software, resolution performance for the all the run modules was calibrated with a quality score cutoff of QV20 (99% basecalling accuracy or a 1% probability of error) using a sliding window of 20 bases from the 3' end. Therefore, while shorter read length specifications are generated with the KB Basecaller, the calibrations are of higher basecalling accuracy. In this release, we are using the same specifications for 98.5% basecalling accuracy, which is on the conservative end, to keep the specifications uniform.

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

We are upgrading to the Windows 10 operating system. What are the compatible capillary electrophoresis secondary analysis software with Windows 10?

The following capillary electrophoresis secondary analysis software are compatible with Windows 10 (and also Windows 7):

-Sequencing Analysis Software v7
-SeqScape Software v4
-Variant Reporter Software v3
-GeneMapper Software v6
-Minor Variant Finder v1.2
-MicrobeBridge v1.1

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

Where do I get the latest Applied Biosystems Capillary Electrophoresis Software patches?

Please visit our Applied Biosystems Genetic Analyzer Freeware and Patches website: https://www.thermofisher.com/sangerpatches

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