Fragments of DNA production methods

the gene machine

artificially synthesises DNA sequence for the known structure of a protein

what the gene machine is dependent on to make a protein

knowing the structure of the protein

Restiction (endonuclease) enzymes

enzymes that break DNA at recognition sites allowing a desired gene to be cut out of the DNA

Reverse transcriptase

used to change mRNA of a virus into cDNA which can be used to code for a desired protein

first extracted in reverse transcriptase method

cell containing mRNA of gene of interest extracted

isolated from the cell in the gene of interest (RV)

mRNA of the gene of interest is isolated from the cell

how a single-stranded complementary copy of cDNA is formed

mRNA acts as a template

the free nucleotides are joined using Reverse Transcriptase

how cDNA is isolated from mRNA template strand

single stranded cDNA is isolated by hydrolysis of the mRNA with an enzyme

how a double stranded DNA is formed from cDNA in the Reverse Transcriptase method

using cDNA as a template and DNA polymerase

recombinant DNA

extracting fragments of DNA from one organism to another

transgenic organism

organism which has received fragments of DNA from a separate organism

PCR (polymerase chain reaction)

method used in labs to create millions of copies of DNA

function of primers

to indicate to DNA polymerase where to form hydrogen bonds with adjacent nucleotides

function of denaturing DNA strand in PCR

splits DNA into two separate strands to be copied in semiconservative replication

temperature to denature DNA in PCR

90∘C

function of annealing in PCR

decrease temperature to allow primers to join to DNA strands

indicate where polymerase should act on

temperature of annealing

around 50∘C

function of DNA polymerase in PCR

joins on complementary free nucleotides to the primer to create a full copy of the desired DNA strand

optimum temperature for DNA polymerase to act

70∘C

how fast the PCR is in creating copies

very fast increases exponentially