Bacterial DNA Barcoding Project: 16S rRNA Amplification Kit
Use PCR to amplify the 16S ribosomal RNA (rRNA) gene, a commonly used DNA barcode for bacterial identification.
Have you ever wondered what types of bacteria are present in the
soil, on your cell phone, or even on your hands? This kit will allow you
to amplify a bacterial DNA barcode, a short region of DNA that can be
used to identify species of bacteria based on their DNA sequence.
Scientists use small parts of the genome called DNA barcodes to
uniquely identify organisms. With this bacterial barcoding kit, and
starting with your own cultured bacteria, you will be able to PCR
amplify the 16S ribosomal RNA (rRNA) gene, a commonly used DNA barcode.
After confirming the results by gel electrophoresis, you may submit your
PCR products to a third party for DNA sequencing, then perform
bioinformatic analysis to identify the bacteria in your samples down to
the genus or species taxonomic levels.
- Techniques: Micropipetting, DNA extraction, PCR, gel electrophoresis, bioinformatics
- Topics: Biodiversity monitoring, DNA barcoding, species identification
- Level: Advanced high school, college, independent research
- Time required: This is a project-based activity that requires at least three class periods for completion
This is an advanced lab activity with
unknown outcomes and is ideal for independent research projects.
Completion of this activity requires access to a DNA sequencing service.
What is included?
Reagents to perform PCR on 48 bacterial samples and to sequence 24 PCR products:
- 5X EZ-PCR Master Mix
- 16S PCR Primer Mix (27FHT-1492RHT)
- 16S Sequencing Primer (1492RHT)
- Nuclease-Free Water
- Fast DNA Ladder 3
- PCR Strip Tubes
Storage:
- If kept in the freezer, reagents can be stored for 12 months after receipt.
- If kept in the refrigerator, reagents can be stored for 1 month after receipt.
Required equipment:
Gel electrophoresis reagents and plastic tubes are supplied by the user
- Agarose
- Gel electrophoresis buffer (e.g. TBE)
- Fluorescent DNA stain (e.g. SeeGreen™ or GelGreen®)
- 1.7 ml tubes for distributing samples to student groups
- 0.2 ml PCR tubes
The Learning Lab Companion Kit (KT-1510-01) provides sufficient reagents to prepare and run eight gels when using the blueGel™ electrophoresis system, as well as 1.7 ml and 0.2 ml plastic tubes.
Materials for culturing bacteria sold separately
- This lab requires supplies for collecting bacterial samples from the environment and culturing them on agar plates.
- The
Microbiology Plate Pouring Kit (KT-1015-02) provides supplies to sample
and grow bacteria on twenty 90 mm Petri dishes using LB agar as the
substrate.
Protocol download
Saftey Data Sheets
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