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Biology Lab > Lab 10 > Flashcards

Lab 10 Flashcards

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1
Q

what are some important rules to remember when handling a micropipette

A
  • do not allow pipettes to come into contact with corrosive chemicals
  • make sure the volume has been correctly set
  • locking ring must be unlocked before changing volumes
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2
Q

how should hold the micropipette

A

keep it upright to prevent liquids from running inside the shaft

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3
Q

what kinds of tips are used with micropipettes

A

disposable ; tips can be autoclaved

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4
Q

what are some of the main structures on the pipettes

A

tip ejector button
digital volumeter
shaft tip holder
tip

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5
Q

what structure in DNA uses a covalent bond

A

the sugar-phosphate sides of the twisted DNA helix

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6
Q

what structures in DNA use a hydrogen bond to be held together

A

nitrogenous bases

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7
Q

why are the first three letters of the name of the enzyme italicized

A

refers to the bacterium from which the restriction enzyme was isolated

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8
Q

what does the fourth letter refer to

A

strain of bacterium if any

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9
Q

what do the numbers refer to

A

the sequence or order in which the enzymes were discovered

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10
Q

what forms the genetic code

A

the exposed bases

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11
Q

type II restriction enzymes

A

particular enzymes isolated from bacteria that can cleave the DNA of bacteriophages

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12
Q

how are cleavage sites determined

A

the enzymes recognizing palindromes of 5’ to 3’ and 3’ to 5’ sequences from four to several base pairs long

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13
Q

how can fragments produced by cutting double stranded DNA be separated

A

by size in a gel treated with an electric current

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14
Q

what is a palindromic sequence

A

nucleic acid sequence whereby reading in a certain direction (5’ to 3’) on one strand is identical to the sequence in the same direction (5’ to 3’) on the complementary strand.

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15
Q

what are the sticky ends of the type II enzymes

A

overhangs are called sticky ends because they can form base pairs with complementary DNA molecules

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16
Q

what does a restriction enzyme reaction contain

A

DNA to be analyzed
a restriction enzyme
restriction enzyme buffer mix

17
Q

what is usually within a restriction enzyme buffer mix

A
  • buffering agent to maintain constant pH
  • salt to provide the correct ionic strength for the digest
  • MG++ as a cofactor for enzyme activity
18
Q

what is a “unit”

A

amount of enzyme needed to digest 1ug of the bacterial virus (lambda phage) DNA in 1 hour in a 50ul reaction

19
Q

how many units do we usually use in lab

A

10-20 units

20
Q

in the lab, what are treating the bacteriophage lambda DNA with

A

restriction enzymes EcoRI, Hindlll, and BamH1

21
Q

what is gel electrophoresis

A

technique used to separate molecules on their sizes and/or charges

22
Q

what is one of the materials the gel can be made out of

A

agarose

23
Q

what is agarose gel electrophoresis

A

used to separate and analyze DNA and RNA

24
Q

why does agarose gel electrophoresis sperate DNA molecules based only on their size

A

because DNA molecules are uniform in charge per mass

25
Q

which way do DNA molecules migrate

A

towards the anode (positive terminal) because they contain negative charges

26
Q

what is agarose

A
  • polysaccharide
  • purified from red seaweed
  • forms a jelly like structure (gel) that contains holes & DNA molecules move through the holes under an electric field
27
Q

what happens when you use more agarose

A

higher concentration, smaller the holes, and slower the DNA would migrate

28
Q

how can DNA migration be sped up

A

with a higher electrical current

29
Q

what are the two most important results from an agarose gel electrophoresis

A

size and amount of DNA

30
Q

what does the position of the DNA indicate

A

size of molecule
- estimated by comparing to molecular marker loaded on same gel

31
Q

what does the band intensity indicate

A

how much DNA within the band
- typically in ng or ug

32
Q

what is a more qualitative way to analyze the sizes of DNA

A

make a plot
- semi log graph can be used to determine the sizes of DNA

33
Q

what is the speed of migration of a DNA molecule based on

A

its size in kb/bp

34
Q

How many positions would a plunger have

A

3
- rest stop
- 1st stop
- 2nd stop

35
Q

why 37⁰C is chosen as the temperature for enzymatic reactions

A

It’s human body
temperature, usually the optimal temperature of an enzyme

Biology Lab (23 decks)

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