lab practical Flashcards
(19 cards)
bacterial transformation
insertion of gene into organism to change that organism’s trait
plasmid
circular DNA with accessory genes in bacteria
purpose of adding ampicillin to plate
- bacteria w/pglo has amp resistance
- amp kills all bacteria w/o pglo plasmid
purpose of adding arabinose to plate
allows gfp to be expressed (glow) by arabinose binding to araC
genes of pglo: araC
regulator protein (on/off switch)
genes of pglo: gfp
makes e coli glow
genes of pglo: bla
amp resistance
genes of pglo: ori
origin of replication
CaCl2’s function in transformation
Ca+ cation neutralizes phosphate backbone + phospholipid membrane so new DNA can be inserted
caluclating transformation efficiency
- amount spread = total amount of pglo added to tube x fraction spread
- # of colonies / amount spread = transformants / microgram
what plates did bacteria GROW on?
+pGLO AA - had amp resistance
+ pGLO A- had amp resistance
- pGLO blank- no amp added
what plates did bacteria GLOW on?
+pGLO AA- had arabinose to turn on
Bradford reagent color before/after
red-brown to blue
why does Bradford reagent change color?
when reagent interacts w/protein, it becomes blue (more blue = more protein)
how to calculate absorbance if sample diluted beforehand?
figure out concentration or set up proportion (?)
cell lysis
break down of cellular membrane + intracellular components released
protein extraction
extract protein through diff methods + separate it from rest of cell components
lysozymes
enzymes that destroy bacterial cell wall
chromatography
separates proteins in complex mixture
