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Applied Biosystems™ MicroSEQ™ Full Gene 16S rDNA PCR Kit
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Description

The MicroSEQ Full Gene 16S rDNA PCR Kit is the PCR component of the MicroSEQ 16S rDNA Full Gene Identification Kit, which provides an easy-to-use DNA sequence-based method to identify most bacteria.

The MicroSEQ™ Full Gene 16S rDNA PCR Kit is the PCR component of the MicroSEQ™ 16S rDNA Full Gene Identification Kit, which provides an easy-to-use DNA sequence-based method to identify most bacteria. Use the PCR kit to amplify the bacterial 16S rRNA gene, for subsequent sequencing using the MicroSEQ™ Full Gene 16S rDNA Sequencing Kit. This PCR kit does not include a protocol or a Quick Reference Card.

  • Sequence-based measurement ensures you get reproducible results in under 24 hours.
  • Growth requirement-independent measurement enables you to identify bacteria using objective criteria.
  • Universal domain-level primers eliminate up-front optimization.
  • MicroSEQ™ ID Analysis Software ensures accurate species identification.
  • Customizable libraries give you the flexibility to create new libraries and search validated libraries that contain the most frequently encountered organisms.
  • Requires minimal training-similar work flow as the MicroSEQ™ D2 LSU fungal kit and the MicroSeq 500 16S rDNA kit.

The MicroSEQ™ ID System: A Complete, Integrated Solution

Streamlining every step in microbial identification, Applied Biosystems™ MicroSEQ™ System combines the advantages of MicroSEQ™ ID Software with easy PrepMan Ultra sample preparation reagents and protocols, MicroSEQ™ bacterial and fungal application kits, and industry-leading thermal cyclers and sequencing systems. To ensure optimal performance and system-wide integration, we develop and test all components of the system together.

Specifications

Content And Storage • 2X PCR Master Mix (sufficient for 20 amplifications), 3 tubes
• Positive Control DNA, 1 tube
• Negative Control DNA (water), 1 tube
• Protocol and Quick Reference Guide

After first use, store at 2°C to 8°C in a PCR clean room.
Detection Method Primer-probe
GC-Rich PCR Performance Low
Technique SBT (Sequence-Based Typing)
For Use With (Equipment) GeneAmp 9700, Veriti Thermal Cycler
Libraries Customizable, Bacterial Library (> 2000 species)
Product Line MicroSEQ
Product Type Full Gene 16S rDNA PCR Kit
Quantity 1 Kit
Shipping Condition Dry Ice
Software MicroSEQ™ ID Analysis Software
Target 16S rRNA (full gene)
Target Organism Class Bacteria
For Use With (Application) Standard PCR
Sample Type DNA (Genomic)
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Frequently Asked Questions (FAQs)

What primers are supplied with the MicroSEQ Full Gene 16S rDNA PCR Kit?

The MicroSEQ Full Gene 16S rDNA PCR Kit includes universal domain-level primers that are designed to amplify the bacterial 16S rRNA gene.
Where can I get a copy of the free trial version of the MicrobeBridge Software?

A 60-day-free trial of the MicrobeBridge Software can be downloaded here: www.thermofisher.com/order/catalog/product/A29238
Who do I contact for support for MicrobeBridge Software and for MicrobeNet database issues?

For the MicrobeBridge Software, please contact your local Thermo Fisher Scientific Technical Support (https://www.thermofisher.com/us/en/home/technical-resources/contact-us.html) or email abtechsupport@thermofisher.com.

For MicrobeNet, please contact the CDC directly at microbenet@cdc.gov. This account is actively monitored by the CDC MicrobeNet staff.
What are the minimum system requirements for installing the MicrobeBridge Software?

The MicrobeBridge Software has the following recommended hardware requirements: Minimum 1.8 GHz processor, 2 GB RAM, and 20 GB of free hard disk space. MicrobeBridge Software has been tested on Windows 7 32-bit SP1, Windows 7 64-bit SP1, and Windows 10 IoT.
Will I lose any data if I upgrade from the MicrobeBridge Software free trial version to the full version?

No data is lost with the MicrobeBridge Software installation of the full version on top of the free trial version.
Which input file formats can I use with the MicrobeBridge Software?

Only .ab1 files can be used with the MicrobeBridge Software.

Which instruments does the MicrobeBridge Software support?

The MicrobeBridge Software fully supports data outputs from Applied Biosystems 3500, 3500xL, 3130, 3130xl, and SeqStudio genetic analyzer systems.
Do I need a MicrobeNet account to use MicrobeNet?

To use MicrobeNet, you will need to register for a MicrobeNet account. You can register at https://microbenet.cdc.gov/

Does MicrobeNet database provide species level identification?

Yes, MicrobeNet database provides species-level identification.
Is there a list of organisms that can be identified with MicrobeBridge Software?

MicrobeBridge Software does not do bacterial identification. It provides a connection to MicrobeNet which does the bacterial identification.
Can I edit the MicrobeNet software search parameters in MicrobeBridge Software?

No. MicrobeBridge Software does not provide any search parameters which are used by MicrobeNet software.
Can I manually override the changes that the MicrobeBridge Software makes to the contig sequence?

Yes, you can edit the contig sequence generated by the MicrobeBridge software.
Can I export the final contig sequence generated by the MicrobeBridge Software to search in other databases?

Yes, the final contig sequence generated by MicrobeBridge Software can be exported for use in other database searches.
Does MicrobeBridge Software do basecalling?

Yes, MicrobeBridge Software uses Applied Biosystems KB Basecaller v1.4.2.4, by default.
Do I need to sequence with a specific chemistry to use MicrobeBridge Software?

As long as the 16S full gene is covered, a specific chemistry is not needed when using the MicrobeBridge Software.
Do you have recommended protocols, reagents, and primers, to generate MicrobeBridge Software samples?

CDC MicrobeNet has recommended primers which have been configured into the MicrobeBridge Software. We recommend using MicroSEQ 16S Full Gene kit for a complete package solution.
Can sequences generated using the MicroSEQ PCR and sequencing kits be analyzed using MicrobeBridge Software?

Data generated from the MicroSEQ 16S Full Gene Kit package can be analyzed using MicrobeBridge Software.
What is the difference between MicrobeBridge Software and MicroSEQ ID software?

Through a partnership with the Special Bacteriology Reference Laboratory at the CDC, Thermo Fisher Scientific developed the MicrobeBridge Software to enable public health, microbiology, and any other laboratories worldwide to directly access the CDC's MicrobeNet virtual reference database using 16S rRNA sequencing technology.

MicrobeBridge Software provides a streamlined workflow for analyzing Sanger sequencing data (.ab1 data files) generated on CE Sanger sequencing instruments or genetic analyzers. MicrobeBridge Software performs contig assembly, contig editing, primer trimming, data quality check, contig sequence copying, and one-click connection to MicrobeNet for bacterial identification through 16S rRNA BLAST searches.

In contrast, MicroSEQ ID software is part of a complete system designed for pharmaceutical analytics workflows. The system includes secondary analysis software, validated reference libraries for bacterial and fungal identification, links to query public domain databases, creation of custom reference databases, and SAE features that enables 21 CFR Part 11 compliance.
Is MicrobeBridge Software a replacement for the MicroSEQ ID software?

MicrobeBridge Software is not a replacement for the MicroSEQ ID software. MicrobeBridge Software is specifically designed for public health, microbiology, and any other laboratories utilizing the MicrobeNet database. Applied Biosystems MicroSEQ ID Microbial Identification software is designed for use in pharmaceutical analytics laboratories who typically do not use the MicrobeNet database but require other functionalities. However MicroSEQ ID users can install and use MicrobeBridge Software if they wish to do MicrobeNet searches.
Who can benefit from the MicrobeBridge Software?

Public health, microbiology, and any other laboratories conducting disease surveillance or bacterial identification with MicrobeNet can benefit from using the MicrobeBridge Software.
What are the benefits of using the MicrobeBridge Software?

Some of the benefits of using the MicrobeBridge Software Faster are faster turnaround time and lower cost:

Using MicrobeBridge Software, MicrobeNet users are able to efficiently assemble and QC their sequencing data using the CDC recommended primer set for 16S rRNA gene sequencing analysis. Users are able to get to their results quickly and accurately, thereby saving money and time spent assembling, interrogating, and identifying bacteria or pathogens in their data.

Users can connect to MicrobeNet via a single click in the software or export the consensus contig and this in turn improves the bacterial identification workflows in public health and microbiology laboratories using MicrobeNet.

Users are able to connect directly into the CDC MicrobeNet virtual reference database for identifying rare and complex pathogens faster and more accurately. Although any laboratory with an internet connection can access the MicrobeNet database worldwide, MicrobeBridge Software streamlines the CE data and connects to MicrobeNet directly.
What is MicrobeBridge Software?

The Applied Biosystems MicrobeBridge Software is a desktop application which connects .ab1 data files generated on Applied Biosystems 3500 and 3130 Genetic Analyzers and 3730 DNA Analyzers with the Centers for Disease Control and Prevention (CDC) MicrobeNet virtual reference database via internet connection for bacterial identification using 16S rRNA gene sequencing analysis.

MicrobeNet is a web-based reference database for the identification of microbial pathogens through 16S rRNA gene sequence BLAST searches. MicrobeNet contains a database of genetic and phenotypic properties for bacterial identification actively curated by the CDC.

For Research Use Only. Not for use in diagnostic procedures.