A common task facing geneticists is to assay for sequence changes at particular locations in genes. These assays are often looking for changes in the coding exon of genes, and the target sequences are typically amplified using PCR from genomic DNA using a pair of specific primers. In this article, we will show you how to use NCBI Reference Sequences and Primer-BLAST, NCBI’s primer designer and specificity checker, to design a pair of primers that will amplify a single exon (exon 15) of the human breast cancer 1 (BRCA1) gene.
Here are the steps to follow to design primers to amplify exon 15 from human BRCA1:
Submitting sequences to GenBank can seem complicated at first, but starting with a solid foundation in the form of a properly formatted file will make the process go smoothly.
Before submitting sequence data to GenBank, the data must be formatted correctly, the most common file format being FASTA. This post will show you how to create a FASTA file for submitting single- and multiple-nucleotide sequences.
Submitters can upload FASTA-formatted sequence files using NCBI’s stand-alone software Sequin, command line tbl2asn or our web-based submission tool BankIt.
The image below depicts a single sequence in FASTA format. For multiple sequences, such as those of population or phylogenetic studies, environmental samples, and batch sequences of the same gene, create the file using the steps below and put the set of sequences together in a single FASTA file.
Here is how to create the FASTA file: