BIOL240 Chapter Notes - Chapter 9: Wood Block, Multiple Cloning Site, Methyltransferase
Document Summary
1 create your mutants: there are ways to increase the frequency of mutations, exposure to uv light (damages dna, dna repair, chemical mutagens, transposons mobile genetic elements that can disrupt genes. Inoculated a flask then plated bacterial culture in the presence of t1. Leaves overhang of single strands of dna that are sticky and ligate again to another complementary strand: purpose is to restrict/digest incoming bacteriophage dna that may be harmful to host. Ligation: dna ligase combines the overhang of strand a and strand b to make recombined versions, reattaches the phosphate backbone of previously cut dna. Dna being put into the host: have a high copy number results in high yield of extracted plasmid later. Low copy number plasmids are used when minimal expression is needed: plasmids are replicated by host cell dna polymerases, plasmids encode genes that regulate rate of plasmid replication and determine plasmid copy number within cell.
