BME100 s2015:Group15 12pmL4: Difference between revisions
Line 77: | Line 77: | ||
<!-- Include a description of the thermal cycling program below. You can use text, a screen capture, a camera snapshot of the computer screen, or a digital drawing (e.g., using shapes and text boxes in Microsoft Powerpoint) --> | <!-- Include a description of the thermal cycling program below. You can use text, a screen capture, a camera snapshot of the computer screen, or a digital drawing (e.g., using shapes and text boxes in Microsoft Powerpoint) --> | ||
Question 1: | |||
# Template DNA: | |||
# Primers: | |||
# Taq Polymerase: | |||
# Deoxyribonucleotides (dNTP's): | |||
Question 2 and 3: | |||
HEATED LID: 100°C | HEATED LID: 100°C | ||
Line 96: | Line 99: | ||
Question 4: | |||
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. | |||
Revision as of 22:55, 31 March 2015
BME 100 Spring 2015 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | ||||||||||||||||||||||||||||||||
OUR TEAMLAB 4 WRITE-UPProtocolMaterials
PCR Reaction Sample List
OpenPCR program Question 1:
HEATED LID: 100°C INITIAL STEP: 95°C for 2 minutes NUMBER OF CYCLES: 35 Denature at 95°C for 30 seconds Anneal at 57°C for 30 seconds Extend at 72°C for 30 seconds FINAL STEP: 72°C for 2 minutes 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.
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
|