A barcode is a 650 base pair long sequence from the COI region in the mitochondria. There are three basic parts to barcoding. The first is data submission so that all the information relating to a specimen is entered into the Barcode of Life Database (BOLD). The second is the actual lab work where you go from having tissue to the sequencing reactions. The final part of barcoding is the analysis of the data where you edit the sequences and put them onto BOLD.

Data Submission

The first step in DNA barcoding is the data submissions. Before you can do any lab work, you need to create a BOLD project (unless one already exists, in which case you must be added to the project), add the specimen information into BOLD in a BOLD Specimen Spreadsheet, upload the plate or box records to LIMS, and put the plates into Dataserv.

Protocols for Lab Work

Once all the data has been entered to LIMS, BOLD and Dataserv, you can do lab work. The first step is tissue lysis, then extraction is either robotic in BIO or manual in the Science Complex. When you have DNA, you then have to run PCR reactions, e-gel them, put all the steps on LIMS, and then sequence the samples.

Analysis of Data

After you have completed all the data submissions and lab work, you can finally analyze the resulting sequences. The sequences have to be edited with a sequence editing program. There are many different programs, but in the Hanner lab we have SeqScape and Sequencher. Once the sequences have been edited, the sequences and trace files have to be uploaded to BOLD.

Ordering, Mailing, and other Miscellaneous Things to Know

This is a list of miscellaneous things that should help with lab work, such as ordering supplies from various places and how to mail things to people.

Created April 2009

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