Submitting DNA to DNA Sequencing Facility

     DNA creates the basis of life. It determines who we are and our traits. Our DNA distinguishes us from plants, and the sequences of our DNA and a plant's DNA are different. For scientists, it is important to be able to sequence DNA to determine the order of nucleotides and to compare the DNA of different organisms. In many cases, knowing the DNA sequence can help make many discoveries. In order to sequence DNA, you will need a purified DNA template, a primer, and a buffer. There are several DNA sequencing facilities, so choose one (usually, they work through colleges or companies).

     The first thing you need to do is to prepare a 1.7mL centrifuge tube. Add 300ng of the template DNA. In order to calculate the amount in ng, use the concentration of the template. If you do not know the concentration, use the nanodrop to find out. Then, choose the primer that corresponds with the DNA template. Add 1uL of 10uM primer OR 4uL of 2.5uM primer. Finally, add nanopure in order to make a final volume of 12uL. When you are done preparing the tube, label it according the the sequencing facility's instructions, and store it as needed.

     When you get your results back, you may need to BLAST (Basic Local Alignment Search Tool) the sequence you received from the facility. A BLAST will tell you if your gene is from the human genome or the genome of another organism, and if so, what organism. This may be able to help you with your research, or it can be used as a check to see if you have the correct DNA. BLASTs are done through NCBI (National Centre for Biotechnology Information). Since your DNA sequence is of nucleic acids, choose nBLAST on the website (http://blast.ncbi.nlm.nih.gov/Blast.cgi). Enter the sequence you received from the DNA sequencing facility, and select the option that allows you to search all known genomes. The red indicates matches, so the more red, the better. If you scroll down on the BLAST page, you will be able to see which organisms your sequence matches. Save the information on the BLAST page in order to reference them later if needed.