C. Elegans Lab Report

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INTRODUCTION Caenorhabditis elegans, also known as C. elegans, are considered to be a model organism. their short life cycle, well annotated genome, small size , and ease of care make them perfect for a laboratory environment ("WormClassroom"). C. elegans feed off of E. coli and are also hermaphroditic self-fertilizers. A single worm can produce several hundred offspring quickly, which then go on to live for another 2-3 weeks. The life cycle of the C. elegans nematodes can last up to three days from egg to egg-producing adult ("WormClassroom"). C. elegans provide a genetic model for understanding questions of many divers areas of biology. Despite their small size and primitive structure, the worms share many biological properties with other…show more content…
elegans move in a sinusoidal wave pattern, as seen in figure 1 below. But as one can see in figure 2, some worms may curl up into a “u” shape and move in a more circular pattern along their long axis. This is a result of a mutation in the rol-6 gene found in the DNA of the nematodes (Higgins and Hirsh 63-72). This rol-6 mutation is what our experiment will focus on. The worms’ DNA will be amplified using PCR, or polymerase chain reaction. The samples are then sent off to a company that will sequence the DNA according to the Sanger sequencing method. The Sanger sequencing method is, “ a simple and rapid method for determining nucleotide sequences in single-stranded DNA by primed synthesis with DNA polymerase” (Sanger and Coulson 441-446). The DNA fragments will also be separated using gel electrophoresis. Depending on the size of the fragments, the negatively charged DNA will travel through the gel towards a positively charged electrode at the opposite end. The smaller the fragment the faster, and therefore farther, it will migrate. The smaller the molecule the farther it moves through the gel and the more easily it slips through the lattice formed by the buffer solution that is mixed with the gel ("Agarose Gel Electrophoresis of…show more content…
coli. The E. coli served as food for the worms. The worms were kept at room temperature and, a week later, the worms had multiplied to a great number and were nearly starved. the worms were washed off the plate in to a micro centrifuge tube with a small amount of distilled water. The tube was then centrifuged for two minutes at a speed of 2,000 rpm. Using a pipet, the distilled water was removed as carefully as possible so as to not disturb the pellet. a lysis buffer was added to break down the membranes of the cells. The tube was then frozen using liquid nitrogen. Once the tube and its contents had completely frozen Proteinase K was added and then the entire tube was incubated for an hour in a water bath at 65ºC. The tube was then boiled for 10 minutes to inactivate the Proteinase K. Once the tube was removed, the isolated DNA was used in a PCR Reaction. The PCR, or a polymerase chain reaction was prepared by adding template DNA, a forward primer, a reverse primer, water, and what is called a ‘mastermix’. The mastermix consists of Taq polymerase, dNTP’s, and water. The PCR amplifies the DNA. It starts by first denaturing the DNA by heating the sample to 94º-96ºC for a couple minutes until the strands of the target DNA separate. Next, the temperature is lowered to around 50º-60ºC for a few minutes allowing the forward and reverse primers to anneal (base pair) to their complementary sequences.

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