Bio Tech :) Flashcards
What is a probe?
It’s a short single-stranded man made RNA or DNA that is complementary to the gene of interest and its fluorescent so that when visualised the gene of interest can be located
what does PCR do and what is required?
Amplifies DNA
requires: DNA template, DNA primers, DNA nucleotide, taq polymarse (heat resistant enzyme)
Describe the process of PCR
DNA is heated weak bonds break and strands separate.
The mixture is cooled and the primers bind to expose the template strands.
Free nucleotide binds to exposed bases through complementary base pairing. Taq polymerase assists with base pairing and sugar backbone. the process is repeated resulting in amplifying the sample.
Describe gel electrophresis
DNA samples placed in the gel
Power supplies are switched on charging the electrodes and DNA is pulled towards the positive end.
smaller fragments will travel faster and further. the sample is visualized.
Why is Taq needed in PCR?
Taq polymerase is heat resistant and goes through the heating and cooling process without denaturing
What is DNA profiling and what is required?
identification examples crime scenes and paternity test (you are not the father)
It needs DNA sample and needs one from the suspect (dad) and unknown sample., PCR, restriction enzyme s(molecular siccors, cuts DNA leaving unpaired bases called sticky ends), DNA probes.
What is the process of DNA profiling
DNA samples are cut intro different sizes by the restriction enzyme. Then DNA separates through gel electrophoresis. The probes locate specific fragments of DNA and the sample is visualised.
DNA sequencing and what is required
Sequencing is required for PCR and electrophoresis with free fluorescent labelled nucleotides. With terminator bases.
What is the process of DNA sequencing
PCR is run for each base. One tube for A T G C. Heating and cooling. when synthesising the DNA the free nucleotide terminate the strands where they bind randomly producing lengths of fragments.
Gel electrohpresis which gives the DNA
Describe how particular genes can be selected using probes and removed using restriction enzymes.
Probes are complementary to the gene of interest. When DNA strands are separated, the probes can bind to the gene of interest. restriction enzymes cut either side of the gene of interest, isolating the DNA and leaving sticky ends
Describe bacterial transformation
The plasmid is cut with the same restriction enzyme. as the gene of interest
The gene of intrest is encoprated into a bacterial plasmid
DNA is encoporated in the host genome
Decribe a micro injection
inject many copies of the gene of interest into the nucleus of the zygote and hope it gets incorporated.
What is electroporation
An electric pulse is used on a nucleus to increase the membrane permeability where the gene of interest in introduced.
Describe how CRISPR, such as CRISPR Cas9, can be used to edit and/or transfer genes.
The Cas9 protein with guide RNA (complementary to gene of interest). The Cas9 will preform a double stranded cut at the specific location. It either introduces new DNA into the cell or the cell repairs with random free nucleotides and becomes silenced because of errors.
