Quiz 4 - DGS4235 Lab **UV Spectrophotometry, Restriction Enzyme Digestion**

1 mL

Amount of DNA expected when processing HOW MUCH blood can be determined by calculating the relative yield of DNA?

meniscus

where should enzyme be dispensed in order to improve accuracy when setting up Restriction Enzyme digest reactions?

frozen

Enzymes are kept ______ until immediately before use to maintain a high level of acitivty

260/280

what ratio from spectrophotometer gives information on DNA purity?

enzyme

what component is added last when setting up restriction enzyme digests?

50 uL

what is the usual volume for a typical RE digestion reaction?

1 unit

What concentration of Restriction Enzyme is enough to completely digest 1 ug of DNA?

Salt and proper pH

What does the 10X enzyme buffer contribute to the RE digest reactions?

1 hour

Standard Restriction enzyme digestion of genomic DNA can be completed in as little as _____

Concentration

______ = (A260 reading - A320 Reading) x Dilution Factor x 50

Absolute DNA Yield

________ = DNA concentration x total purified sample volume

Relative DNA yield

______ = Absolute DNA yield / volume blood used

DNA purity

______ = (A260 reading - A320 reading) / (A280 reading - A320 reading)

nanodrop

UV spectrophotometer developed for microvolume measurements

No dilution needed