BME100 s2014:W Group13 L4
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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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LAB 1 WRITE-UP
Initial Machine Testing
1. The DNA is seperated into two strands from its normal double helix shape in order to make it readable by heating the DNA up to 95 degrees celcius in the PCR heat block.
2. Primers (strands of DNA which contain the complementary sequence to the start and end points of the strand we want) bind to the strands. This is facilitated by the heat block dropping to a temperature of 57 degrees celcius.
3. The DNA polymerase attaches to the strand at the starting point primer and creates a complementary strand until it falls off of the end of the DNA strand. The PCR machine facilitates this by heating the heat block back up to 72 degrees celcius.
The machine does thirty five rounds of this, which creates over a billion fragments of the target sequence.
When we unplugged (part 3) from (part 6), the LED display on the machine turned off.
When we unplugged the white wire that connects (part 6) to (part 2), the machine's temperature dropped significantly.
The date we tested the machine on was March 19, 2014. We accidentally unplugged the machine and it messed it up. We ran it again and it worked fine.
Thermal Cycler Program
PCR Reaction Mix
Research and Development
PCR - The Underlying Technology
Question 1) The function of each PCR reactions are as follows. The template DNA is the base of the replication of the DNA, without it the DNA cannot be amplified by itself so it is an essential part of the PCR reaction. The primers are used to bind the DNA at the origin to start the replications. The Taq Polymerase is used to create a complementary strand of the DNA strands. The dNTP's (Deoxyribonucleotides) are the building blocks of the ordinary strands.
Question 2) Thermal Cycling The Denature step involves seperating the two DNA strands from each other at the temperature of 95 degrees celsius The Anneal step involves the primers being attached to the DNA strands The extend step involves the polymerase attaching to the strand of the end of the primer The final step involves the polymerase creating a complementary strand of the DNA
Question 3) Each of the DNA nucleotides are paired with only one type, the Adenine pairs with the Thymine and vice versa while the Guanine pairs with the Cytosine and vice versa.