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13 Cards in this Set
- Front
- Back
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What enzyme can digest DNA?
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Restriction endonuclease
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What enzyme can convert RNA to DNA?
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Reverse transcriptase
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What does DNA ligase do?
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Catalyses formation of phosphodiester bonds so facilitates joining of DNA
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Describe how you would encorporate a gene into a bacterial cell
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Plasmid removed from bacterium, DNA containing gene of interest removed from cell. Restriction endonuclease used to create sticky ends on both. Plasmid may take up gene (recombinant plasmids) or may not. DNA ligase forms phosphodiester bonds between sticky ends. Plasmid may be taken up by bacteria = transformed. Growth of bacteria on selective media
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How is complementary DNA produced?
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Genomic DNA is transcripted and spliced into mRNA in cell. mRNA removed from cell, reverse transcriptase converts RNA to DNA (cDNA) which can then be cloned into plasmid.
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Glycosylation of proteins occurs in eukaryotes or prokaryotes.
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Eukaryotes
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What are 3 purposes of glycosylation of proteins?
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Proteins can fold correctly
More stable Cell-cell adhesion |
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Briefly describe PCR
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Amplify a sequence of DNA.
(Denature) Heat to 95degrees C, separates strands of DNA (Anneal) Cool to 50-65dC to allow primers to anneal. (Elongate) Taq polymerase added at 72dC which polymerases complementary DNA to strand. Repeat |
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What is dideoxy chain termination?
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C3 usually has OH, dideoxynucleotide has C3 with just H which terminates chain elongation
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Briefly describe Sanger sequencing
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Four reaction mixtures set up: DNA, primer, DNA polymerase, nucleotides and dideoxynucleotides of A,C,G,T.
DNA run on electrophoresis gel, small portions move further towards positive end. |
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Briefly describe Southern Blotting
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DNA separated on agarose gel by electrophoresis.
DNA transferred to nitrocellulose nylon membrane (Blot). Radiolabelled probe added, complementary to target sequence. Membrane covered with x-ray film to visualise where target sequence is |
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What is a microarray?
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An arrangement of DNA probes on glass slides or nylon membranes
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What is Restriction Fragment Length Polymorphism analysis?
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Separate different DNA strands by the variation between different strands. Restriction enzyme used to cut DNA and then run on gel electrophoresis gel
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