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recombinant DNA
DNA from multiple sources combined
uses plasmids and restriction enzymes
plasmid features
selectable marker (antibiotic resistance)
reporter gene
multiple cloning site (MCS)
polymerase chain rxn (PCR)
amplifies DNA
denaturation
annealing
extension
uses Taq polymerase
PCR applications
forensics, mutation detection, and gene expression (RT-PCR, qPCR)
gel electrophoresis
separates DNA
southern blot analysis
DNA
northern blot analysis
RNA
FISH analysis
fluorescent detection
sanger sequencing
chain termination
NGS
high-throughput sequencing
TGS
long reads
transgenic organism genetic engineering
added gene
knockout genetic engineering
gene removed
CRISPR-Cas9
guide RNa directs Cas9 to DNA
cuts DNA at specific sequence (PAM required)
CRISPR applications
disease treatement (sickle cell) and genome editing
risks of genetic engineering
off-target mutations