lab practical Flashcards
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 (?)
