Today we had our first lab (what an experience!) in which students started extracting DNA from their own blood. We had the assistance of Dr. Lisa Walden and her students from ONU's Clinical Laboratory Science, who very kindly offered to do the phlebotomies necessary to obtain the samples.
The exercise took longer than expected, so we didn't finish the process, but the samples were frozen after the cell lysis step and will be ready to continue with the addition of phenol-cholorform-isoamyl alcohol (PCI) in the next lab session.
Tomorrow: Restriction Enzyme Analysis (RED) of lambda-DNA.
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Friday, September 10, 2010
Module 1, Lab 02 - Restriction Enzyme Digestion (RED) of Lambda DNA
A micrograph of multiple bacteriopages
Restriction enzymes are one of the most basic and important tools in molecular biology. They evolved in bacteria to attack and cut (cleave) foreign DNA, mostly from bacteriophages (viruses that "eat" bacteria). But hey have been isolated to be used in the lab, and are useful to cut any kind of DNA, not just viral.
Cleaving DNA is the first step in any technique that involves recombinant DNA technology. There are techniques that use special enzymes to paste (ligate) different fragments of DNA. For instance a gene can be ligated into a plasmid that can be inserted into bacteria to make many copies of it via bacterial reproduction (cloning), something we will do in a few weeks.
Today we used lambda DNA (DNA from the common lambda bacteriophage) as the substrate to be cleaved with three different restriction enzymes: EcoRI, HindIII, and PstI.
Cleaving DNA is the first step in any technique that involves recombinant DNA technology. There are techniques that use special enzymes to paste (ligate) different fragments of DNA. For instance a gene can be ligated into a plasmid that can be inserted into bacteria to make many copies of it via bacterial reproduction (cloning), something we will do in a few weeks.
Today we used lambda DNA (DNA from the common lambda bacteriophage) as the substrate to be cleaved with three different restriction enzymes: EcoRI, HindIII, and PstI.
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