Ion 16S™ Metagenomics Kit - FAQs

View additional product information for Ion 16S™ Metagenomics Kit - FAQs (A26216)

16 product FAQs found

What is the Ion 16S Metagenomics Kit designed for?

The Ion 16S Metagenomics Kit is designed for rapid analyses of polybacterial samples using Ion Torrent Sequencing technology. The kit includes two primer sets that selectively amplify the corresponding hypervariable regions of the 16S region in bacteria:

- Primer set V2-4-8
- Primer set V3-6, 7-9
Sequence the amplified fragments on the Ion PGM System and analyze the results with the Ion Reporter software Ion 16S Metagenomics Kit analyses module. The combination of the two primer pools allows for sequence-based identification of a broad range of bacteria within a mixed population.

I have run out of Environmental Master Mix v2.0 from the Ion 16S Metagenomics Kit (Cat. No. A26216). Do you offer this as a standalone item?

Yes. You can purchase the TaqMan Environmental Master Mix 2.0 (Cat. No. 4396838).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center

How many reactions can be performed with the Ion 16S Metagenomics Kit (Cat. No. A26216)?

For each sample, the protocol involves two PCR reactions, one with primer set V2-4-8 and one with primer set V3-6.7. However, the kit contains sufficient master mix for 100 reactions and each of the two primer pools has sufficient volume for 100 reactions. Thus, the kit contains sufficient reagents for 100 reactions/50 samples.
Note: 100 reactions is dependent upon on the master mix, which is the limiting reagent. As far as primers are concerned, there is sufficient volume to process another 50 samples.

How many PCR reactions can I perform using the 2X Environmental Master Mix within the Ion 16S Metagenomics Kit (Cat. No. A26216)?

It is sufficient for 100 PCR reactions.

Can I use the Ion 540 Kit - Chef to prepare samples from the Ion 16S Metagenomics Kit?

No, the Ion 16S Metagenomics Kit utilizes 400 bp reads, so it is incompatible with the Ion 540 Kit - Chef, which only yields 200 bp reads.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

If I accidentally use the Ion Library TaqMan Quantitation Kit instead of the Ion Universal Library Quantitation Kit with my Ion 16S Metagenomics libraries, what might happen?

This may result in a lower observed library yield as determined by qPCR.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How long does it take to analyze Ion 16S Metagenomics data in Ion Reporter?

The data can be analyzed in less than 1 hour.

What databases are used by the Ion 16S Metagenomics Ion Reporter workflow?

The 16S Metagonics Ion Reporter workflow uses the MicroSeq ID and GreenGenes databases.

How do I analyze the data generated by the Ion 16S Metagenomics Kit?

Use the 16S Metagenomics workflow in Ion Reporter.

How long does it take to complete the Ion 16S Metagenomics Kit workflow?

The workflow, which consists of library generation, template prep, sequencing, and analysis, takes 2 days.

What kit should I use to quantitate library input generated by the Ion 16S Metagenomics Kit?

We recommend using the Ion Universal Library Quantitation Kit, Cat. No. A26217. It is compatible with the uracil-containing amplicons generated by the Ion 16S Metagenomics Kit.

What control DNA is included in the Ion 16S Metagenomics Kit?

The Ion 16S Metagenomics Kit includes E. coli control DNA (strain K-12).

Is the TaqMan Environmental Master Mix 2.0 that is used in the Ion 16S Metagenomics Kit available separately?

Yes, the TaqMan Environmental Master Mix 2.0 is available separately as Cat. No. 4396838.

What PCR mastermix is used by the Ion 16S Metagenomics Kit?

The Ion 16S Metagenomics Kit uses TaqMan Environmental Master Mix 2.0.

What variable regions are covered by the Ion 16S Metagenomics Kit?

The Ion 16S Metagenomics Kit uses two primer sets to selectively amplify the corresponding hypervariable regions of the 16S rDNA gene in bacterial samples:

- 16S Primer Set V2-4-8
- 16S Primer Set V3-6,7-9

What is 16S ribosomal RNA (rRNA)?

16S is a component of the 30S small subunit of a prokaryotic ribosome. It is a ˜1500 bp region that can be used to identify and classify microbes.