Lab Chapter 4: DNA Fingerprinting

Question 1

What are recognition sites?

Answer 1

Groups of enzymes in bacteria that break/hydrolyze the sugar-phosphate backbone at specific recognition sites

Question 2

What are restriction endonucleases?

Answer 2

a break in sugar-phosphate backbone of both strands of a duplex of DNA causes molecule to become fractured in two pieces. The enzymes that do those are restriction endonucleases

Question 3

What is EcoRI?

Answer 3

a restriction endonuclease that is used routinely in laboratories around the world.

Question 4

What are overhangs/sticky ends?

Answer 4

when the restriction enzyme generates fragments that have single-stranded ends

Question 5

Describe the process of pipetting properly

Answer 5

a. Inspect which size and then recheck
b. Dial up desired volume then recheck
c. load proper size tip, do you need a yellow or blue
d. Place thumb on top of control button

Question 6

Describe process of drawing sample

Answer 6

a. press down to first stop and pause
b. place tip onto sample
c. allow control button to slide slowly to full height
d. slowly pull tip out of sample

Question 7

Describe process of releasing sample

Answer 7

a. insert tip containing sample into desired vessel
b. slowly press control button past first stop and continue to second stroke (blow-out) point
c. holding control button at blowout point, remove tip along wall of vessel, and out of vessel
d. slowly release control button
e. eject tips into proper container

Question 8

True or False: You should always turn the pipet upside down when in use

Answer 8

false: NEVER turn pipet upside down when in use as you will lose your sample

Question 9

True or False: when in doubt as to whether a tip is clean or has been contaminated, change the tip

Answer 9

True

Question 10

___ ul of enzyme mix shuld be put into each of the colored tubes, using a fresh tip for each sample, which each contain ___ul of DNA

Answer 10

10uL

Question 11

The total reaction volume after adding the enzyme mix to the DNA samples for each mix is

Answer 11

20uL

Question 12

After adding the enzyme mix to the DNA samples, the volume should be ___

Answer 12

mixed by flicking and tapping, then pulse spin in microcentrifuge

Question 13

When incubating, the samples should be placed at ___ C for ___ minutes, then ___

Answer 13

37*C
45
then place in water bath to maintain temperature and pulse spin again in microcentrifuge

Question 14

After pulse spinning in microcentrifuge, place group’s tubes ____

Answer 14

into freezer box on front bench