BME100 s2015:Group15 12pmL4: Difference between revisions
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Question 1: | Question 1: | ||
# Template DNA: | # Template DNA: The strand of DNA that is used as a model for replication or the DNA waiting to be copied during the PCR experiment. | ||
# Primers: | # Primers: Sequences of nucleotides that bind to specific locations on the template DNA. Primers come in pairs so they can bind on each strand of the DNA. | ||
# Taq Polymerase: | # Taq Polymerase: The enzymes that are actually being replicated based upon the primers that were combined on the template DNA. They bind the first primers and then add necessary nucleotides that complete the original strand as it moves down the template DNA | ||
# Deoxyribonucleotides (dNTP's): | # Deoxyribonucleotides (dNTP's): The monomers that are needed to complete a strand of DNA. They are responsible for the replication of the template DNA and are what the taq polymerase connect with form the strand. | ||
Question 2 and 3: | Question 2 and 3: | ||
HEATED LID: 100°C | #HEATED LID: 100°C | ||
INITIAL STEP: 95°C for 2 minutes | #INITIAL STEP: 95°C for 2 minutes | ||
NUMBER OF CYCLES: 35 | #NUMBER OF CYCLES: 35 | ||
Denature at 95°C for 30 seconds | #Denature at 95°C for 30 seconds | ||
Anneal at 57°C for 30 seconds | #Anneal at 57°C for 30 seconds | ||
Extend at 72°C for 30 seconds | #Extend at 72°C for 30 seconds | ||
FINAL STEP: 72°C for 2 minutes | #FINAL STEP: 72°C for 2 minutes | ||
FINAL HOLD: 4°C | #FINAL HOLD: 4°C | ||
First the sample from the template DNA is taken. The template DNA is the original DNA segment that is to be copied. The dNTP’s are the nucleotides of the template DNA. During step one, the PCR tube is kept at 95 degrees celsius and the double helix separates and creates two separate strands. Then the PCR tube is cooled down to 57 degrees and the primers attach to their target sites. “Primers attach to sites on the DNA strand that are on either end of the segment that you want to copy.” The temperature is then increases to 72 degrees and the DNA polymerase is then activated and finds the necessary nucleotide and attaches it to the corresponding strand. Taq polymerase reads the code of a strand and attaches a corresponding nucleotide to create a copy of the DNA. Adeine binds to Thymine, and Cytosine binds to Guanine. The tube is then kept at 72 degrees until the target strand is fully replicated. | First the sample from the template DNA is taken. The template DNA is the original DNA segment that is to be copied. The dNTP’s are the nucleotides of the template DNA. During step one, the PCR tube is kept at 95 degrees celsius and the double helix separates and creates two separate strands. Then the PCR tube is cooled down to 57 degrees and the primers attach to their target sites. “Primers attach to sites on the DNA strand that are on either end of the segment that you want to copy.” The temperature is then increases to 72 degrees and the DNA polymerase is then activated and finds the necessary nucleotide and attaches it to the corresponding strand. Taq polymerase reads the code of a strand and attaches a corresponding nucleotide to create a copy of the DNA. Adeine binds to Thymine, and Cytosine binds to Guanine. The tube is then kept at 72 degrees until the target strand is fully replicated. |
Latest revision as of 23:23, 31 March 2015
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OUR TEAMLAB 4 WRITE-UPProtocolMaterials
PCR Reaction Sample List
OpenPCR program Question 1:
First the sample from the template DNA is taken. The template DNA is the original DNA segment that is to be copied. The dNTP’s are the nucleotides of the template DNA. During step one, the PCR tube is kept at 95 degrees celsius and the double helix separates and creates two separate strands. Then the PCR tube is cooled down to 57 degrees and the primers attach to their target sites. “Primers attach to sites on the DNA strand that are on either end of the segment that you want to copy.” The temperature is then increases to 72 degrees and the DNA polymerase is then activated and finds the necessary nucleotide and attaches it to the corresponding strand. Taq polymerase reads the code of a strand and attaches a corresponding nucleotide to create a copy of the DNA. Adeine binds to Thymine, and Cytosine binds to Guanine. The tube is then kept at 72 degrees until the target strand is fully replicated.
Base pairing happens during the annealing period and the extension period. As the system cools off, the target DNA are blinded with the primers so that the replication process can occur. This is part of the annealing process. During the extension stage of the thermal cycle, when the temperature is elevated, is when the DNA comes into existence. In this stage two taq polymerase match with base primers created in the previous stage, which in turn create a new nucleotide stand that sequences DNA. These are the two steps that make DNA replication possible in a PCR model experiment.
Research and DevelopmentPCR - The Underlying Technology
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