BME100 f2014:Group30 L4
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Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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LAB 4 WRITE-UP
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, and Extend at 72°C for 30 seconds
FINAL STEP: 72°C for 2 minutes
FINAL HOLD: 4°C
Research and Development
PCR - The Underlying Technology
Question 1: Functions of PCR Components:
The template DNA is the DNA sequence that is copied during the process of mRNA production. The base pairs of the template strand are then copied by DNA primers, as the primers attach to the desired DNA sequences to be copied. Taq polymerase binds to the DNA primers and adds corresponding nucleotides to the template strand in order to complete the second strand of the DNA. Since DNA is made up of deoxyribunucleotides, the deoxyribonucleotides are used to hold the double stranded DNA molecule together.
The initial step is to raise the temperature to near boiling. Causing the double-stranded DNA to separate, or denature, into single strands. When the temperature is decreased, DNA sequences known as primers bind or anneal to complementary matches on the target DNA sequence. The primers bracket the target sequence to be copied. At a slightly higher temperature the enzyme taqpolymerase binds to the prime sequences and adds nucleotides to extend the second strand. This completes the first cycle. In subsequent cycles, the process of denaturing, annealing, and extending are repeated to make additional DNA copies. After three cycles, the target sequence, defined by the primers, begins to accumulate. After 30 cycles, as many as a billion copies of the target sequence are produced from a single starting molecule.
Question 3: Corresponding Base Pairs:
The following nucleotide base pairs bind or anneal to each other during the process of of DNA replication during the PCR reaction.
Adenine (A) - Thymine (T); Thymine (T)-Adenine (A); Cytosine (C)- Guanine (G); Guanine (G)- Cytosine (C)