SA2

The use of technology to modify organisms to make them more useful to humans

Modifying a DNA sequence by removing an unwanted gene or inserting a desired gene Modifying the genotype to produce a desired phenotype

A group of proteins found in bacteria Highly specific - ALWAYS cut at restriction sites *at the backbone

To carry out Restriction digestion Cut DNA by breaking phosphodiester bonds

Pieces of DNA cut by restriction enzymes

When all the bases of cut DNA remain paired

When the bases of cut DNA our unpaired

To join pieces of DNA back together By forming phosphodiester bonds

Any DNA sequence that contains DNA from another source

To amplify DNA To make many copies of a DNA sequence *number of DNA doubled after each round

• Denaturation • Annealing • Extension

Sample heated Hydrogen bonds broken Double-stranded DNA converted to single-stranded DNA

Sample cooled Hydrogen bonds form between single-stranded DNA and primers Primers anneal to single-stranded DNA

Sample heated Taq polymerase binds to primers and adds DNA nucleotides Form double-stranded DNA

To separate DNA based on fragment length Used to confirm that the correct gene/ sequence has been amplified with PCR or for DNA profiling

Cuts out specific, whole sequences of DNA Locates target sequence inside cells and deactivates them by cutting them Can remove unwanted sequences or add desired sequences *permanent

Sequences that are recognized and cut out by enzymes and proteins coded for by the Cas genes Located between CRISPR sequences

Codes for proteins and enzymes that recognize and cut out DNA sequences that match spacer DNA

Where spacer is located Sequences where we insert spacer DNA

Any organism that has gained a new gene or allele through artificial means

Making copies of functional genes that can produce proteins • Producing specific proteins

• Extract plasmid • Cut out desired gene from chromosome and cut plasmid with same restriction enzyme • Mix desired gene with cut plasmids and DNA ligase - produce recombinant plasmids • Mix recombinant plasmid with bacterial culture - so plasmids absorbed into bacterial cells

Plasmid containing the new gene

The process of absorbing the recombinant plasmid Plasmids taken up by bacteria