PCR and Sequencing
When tissue is sent to Smithsonian, Autogen robotics using a modified CTAB/Chloroform extraction protocol is utilized. DNA samples are transferred to DNA barcode tubes with unique extraction barcode numbers.
DNA extractions sent to Smithsonian, will likewise be transferred to DNA barcode tubes with unique extraction barcode number.
Gene regions to be analyzed:
PCR is conducted via a standard protocol – with a maximum of 3 separate attempts to recover amplicons. Successful PCR are collected and sequenced on an ABI 3700 96-well capillary sequencer.
Depositing Sequences in NMNH Barcode Database and CTFS Database
Sequences, including FASTA sequence files as well as links to raw data (chromatograms), are supported for each sample, along with all information originally collected via the standard sample template (see # 1).
The on-line searchable database will be developed with barcoding informatics tools for identification of sequences against the NMNH and CTFS database.