Oct 8, 2009
-In order to understand the basic structure of DNA it is important to understand
the prevalent primary and secondary structure of it.
-Protein is the main component of DNA both in terms of incorporation and in the
construction of the molecule; the first protein to be sequenced is insulin.
-Dideoxy-nucleotide chain termination method is one of the oldest ways to
sequence a protein, but it is considered old and inefficient compared to modern
ways of sequencing (7.12 Singer & Burk)
-The L isoform of amino acids are the ones that are used in proteins, not D.
-In DNA replication there needs to be a template strand to indicate the specificity
of which amino acid to insert along with other chemicals.
-If you want to stop the synthesis process you can use ddATP since it is missing
3’OH, because there in no OH to attach the next nucleotide to.
-When reading the sequence from a gel, you start at the bottom and move
upwards, each of the nucleotides will occupy a different lane.
How to Get a ss Template
-The first step is to find an organism that can make a ss form of DNA and then
infect with a bacteria phage (M-13 in this case)
-The phage will exist in a ss form but will reproduce in a ds replicative form, the
resulting progeny are also in a ss form.
-Use a universal primer that can then bond to the ds replicative form of the
phage and have it create an recombinant molecule (universal primers are just
-The phage will create a hybrid onto the primer and replicate the sequence of your
choosing and then revert back to the ss form.