In this lab, we will be using PCR to examine our own DNA sequence in a test tube. We will be looking for a particular piece of DNA that is present in most people, but not all people. We will be determining this through PCR, which has begun transforming molecular biology into a multidisciplinary research field. On the first day we will be extracting DNA from our cheek cells. We will break open the cell membranes by putting the samples into a water bath, then we will use instagene matrix beads to kill DNase. On day two we will perform PCR in three steps, as shown below....
In this lab PCR has three main steps. The first is denaturation, the second is an annealing step, and the third, an extension step. In the first step, the DNA seperates into two single stranded molecules. The templates must be sperated before the polymerase can generate a copy.
During the annealing step the oligonucleotide primers find their complementary sequenceds on two single stranded template strands of DNA. After they are annealed they can be used as primers for the Taq DNA polymerase. In extension Taq DNA polymerase's job is too add nucleotides (A,T, G and C) to the primer to create a complementary copy of the DNA template.
After running through this, on day three we will use gel eletrophoresis to diagram our results.
No comments:
Post a Comment