In this experiment, three strains of tetracycline resistant E. Coli were tested. First, the bacteria were diluted via serial dilution in order to more accurately count and quantify the bacteria. To ensure that the three different strains of bacteria are not cross contaminated, label everything either Farm A, Farm B, or Farm C depending on what is being tested. There are six agar plates for each farm, three of the plates are tetracycline plates (which are labeled) and three are unlabeled. For the three labeled plates, mark each with their dilution which will be: 10-2 ,10-4, or 10-6; do the same for the unlabeled plates. Next, the dilutions were made using sterile technique so that unwanted bacteria is not introduced to the experiment. Take
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PCR uses DNA primers to bind and recognize the specific regions of DNA that will be amplified. PCR allows scientists to target a specific gene of interest in order to study. To begin PCR analysis, start by taking six tubes (labeled 1-6) that contain PCR reaction mix. Three of the tubes have primers, for each of the tubes with primers touch a pipette tip to one of the colonies that grew on the tetracycline labeled plate and dip into the tube containing the primer. Change the pipette tip and do the same for the other tubes and plates. Bacteria from one plate is put into one tube. Do not mix the samples of bacteria. The other three tubes do not have a primer, put the bacteria from the unlabeled control plates into the tubes without a primer in the same fashion as above. Put the three tubes in the PCR machine and let …show more content…
Gel electrophoresis allows the products of the PCR reactions to be analyzed in order to determine the identity of the tetracycline resistance plasmid. First, in order to see the DNA fragments, move across the gel, add 2 uL of bromopenol blue tracking dye to each of the tubes. In the first well add 10 uL of the PCR DNA ladder as a comparison. Carefully load 15 uL from each of the samples into the wells following the chart below. After the samples are loaded, place the lid onto the gel rig, insert electrode into power supply set at 160 volts. For the best results, let the gel run until the blue DNA fragments are half way up the
PCR uses DNA primers to bind and recognize the specific regions of DNA that will be amplified. PCR allows scientists to target a specific gene of interest in order to study. To begin PCR analysis, start by taking six tubes (labeled 1-6) that contain PCR reaction mix. Three of the tubes have primers, for each of the tubes with primers touch a pipette tip to one of the colonies that grew on the tetracycline labeled plate and dip into the tube containing the primer. Change the pipette tip and do the same for the other tubes and plates. Bacteria from one plate is put into one tube. Do not mix the samples of bacteria. The other three tubes do not have a primer, put the bacteria from the unlabeled control plates into the tubes without a primer in the same fashion as above. Put the three tubes in the PCR machine and let …show more content…
Gel electrophoresis allows the products of the PCR reactions to be analyzed in order to determine the identity of the tetracycline resistance plasmid. First, in order to see the DNA fragments, move across the gel, add 2 uL of bromopenol blue tracking dye to each of the tubes. In the first well add 10 uL of the PCR DNA ladder as a comparison. Carefully load 15 uL from each of the samples into the wells following the chart below. After the samples are loaded, place the lid onto the gel rig, insert electrode into power supply set at 160 volts. For the best results, let the gel run until the blue DNA fragments are half way up the