Unit Five

DNA Isolation and Cloning of
New 16S Community Libraries

CONCEPTS
Biology
-Topoisomerases and ligases in cloning
-The lac operon, genetics and applications
-Antibiotics, selection, and plasmids
-Transformation and heat shock response

Chemistry
-Organic extractions
-Effects of pH, Mg, and salt on PCR

REVISITING PRIOR METHODS
-DNA isolation (Genomic & plasmid mini-prep)
-PCR-for cloning and detection
-Gel Electrophoresis
-Restriction Enzymes (to detect clone insert)

SPECIAL EQUIPMENT
-Epicenter Master-Amp PCR Buffers
-TaKaRa Taq Polymerase
-Invitrogen's TOPO-TA Cloning Kit

"Red Layer" hot spring community.

Phenol/Chloroform extraction reagents.