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Impact of science and technology I
Terms in this set (34)
What does PCR stand for?
Polymerse Chain Reaction
In PCR what is copied?
A section of DNA
The primers used in PCR must have an exact match where in the chain but they don't necessarily need a perfect match for all the other bases?
At the 3' end of the primer
In PCR the new strand is elongated in which direction?
What is a duplex?
Double stranded polynucleotide
The duplex created from the first two steps of PCR is denatured by heating to what temperature?
When the duplex is heated to 95 degrees in PCR, what happens to the two strands?
Once the two strands of the duplex have separated after being heated to 95 degrees in PCR, the mixture is cooled to what temperature which causes what to happen?
Primers to anneal
The new strands that are made from the primers annealing after being cooled to 52 degrees in PCR, what happens to them?
They are elongated by the primers
Does PCR only happen once per DNA fragment?
No the cycle repeats so that you have a large quantity of copied DNA
The product length of PCR is determined by what?
The distance between the two annealing sites of the primers
Name 2 ways that false products can be produced in PCR:
Primers bind to each other (primer dimers)
Primers bind at different sites so product length is different
When PCR is finished what technology are the products put through in order to compare lengths to a band of specific lengths to work out which is the desired product?
For gel electrophoresis what two types of control are required?
-ve and +ve
What is the -ve control for PCR gel electrophoresis?
Shows what the primer dimer will look like
What is the +ve control for PCR gel gel electrophoresis?
Shows what the desired sequence will look like but at a smaller quantity than it is expected to be produced
End-point PCR has just been described, why is it not the best method of PCR?
The amount of product produced does not relate very well to the amount of templates
What is the better method of PCR called that follows PCR over time?
Real time PCR
Is the final yield of PCR a measure of how much template you originally had?
Give four examples of what PCR can identify:
Changes in gene expression
During PCR the ... sequence is amplified by a pair of ... that ... to specific sequences on ... strands of a section of DNA, amplification only occurs between those ... primer sites
What does STR stand for?
short tandem repeats
STR profiling identifies individuals by what method?
Why do STR's allow you to work out the identities and associations of people?
Every person has a different set of short tandem repeats
What percentage of the genome has no clear function?
The majority of the different between the DNA of individuals is in what is described as junk DNA, what is this junk DNA?
Repetitive sequences that vary in the number of repeats
What can STR's be used to follow?
How can you work out whether a child is yours by using STR?
If a child is their parents they will have half their alleles for STR's from one parent and half from the other
Sanger sequencing is the ... sequence of many long molecules
What happens during Sanger sequencing?
The polymerisation of DNA is interrupted at specific bases because as well as normal dNTPs, there are a smaller number of di-deoxy NTPs with different coloured fluorescent dyes attached which are chain terminating so when a ddNTP is incorporated into the sequence we know the end base (by the colour), eventually a ddNTP will be incorporated into every base position so you will know the entire sequence when the results are out through a chromatogram
What are 5 benefits for next generation sequencing over Sanger sequencing?
Uses shorter lengths
Both Sanger sequencing and next generation sequencing can use DNA that has been made from an RNA template as well as original DNA -true/false?
Why is the breakthrough of cheap whole genome sequencing such an amazing diagnostic tool?
A patients' entire genome is known so doctors can work out all the variations in genes that you have so that your drugs and treatments can be tailored to you specifically as they know how your body will react more precisely
How could whole genome sequencing affect patients taking more responsibility for their own health?
If they know they have a mutation that increases their risk for a disease, they are more likely to avoid other risk factors for that disease
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