Topic #14 ( molecular biology)

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1

To extract DNA from cells, DNA must be ____ from the other types of biological molecules in the cells.

separated

2

3 easy steps to extract the DNA:

Detergent eNzymes Alcohol

3

Why add detergent?

To get the DNA out of cells you need to rupture both the ____ membrane and the _____ membrane.

cell, nuclear

4

Why add detergent?

Cell membranes and nuclear membranes consist primarily of

lipids

5

Why add detergent?

Detergents, like all soaps, break up

lipids

6

Why add detergent?

This is why you use detergents to remove fats (which are_____) from dirty dishes.

lipids

7

Why add detergent?

After addition of detergents the membranes are _______ and the DNA is ________ into the solution.

disrupted, released

8

Why add enzymes?

The nucleus of each of your cells contains _____ long strands of DNA with all the instructions to make your entire body.

multiple

9

Why add enzymes?

To fit all of this DNA inside a tiny cell nucleus, the DNA is wrapped tightly around proteins, including

histones

10

Why add enzymes?

The enzyme in meat tenderizer is a _____, which is an enzyme that ____ proteins into small pieces.

protease, cuts

11

Why add enzymes?

As this enzyme (protease) cuts up the proteins, the DNA will ____ and _____ from the proteins.

unwind, separate

12

Why add alcohol?

Cold alcohol _____ the solubility of DNA.

reduces

13

Why add alcohol?

When cold alcohol is poured into an _____ solution containing DNA, the DNA ______ out into the _____ layer,

aqueous, precipitates, alcohol

14

Why add alcohol?

When cold alcohol is poured into an aqueous solution containing DNA, the DNA precipitates out into the alcohol layer, while the____ and_____ stay in the solution.

lipids,proteins

15

A powerful molecular tool using DNA is a _______ technique using _____enzymes that are___ from bacteria.

DNA fingerprinting, restriction, isolated

16

Restriction enzymes recognize specific ____sequences and ____ these sequences.

DNA, CUT

17

With this technique (DNA fingerprinting), large genomic _____ can be cut into smaller _____, or_______

chromosomes, pieces, restriction fragments

18

Because every person's DNA is unique (with the exception of_______), looking at various pieces of DNA from different people would allow for discrimination of DNA between individuals, or rather, a kind of DNA fingerprint.

identical twins

19

Once DNA is cut a researcher must be able to _______ the DNA.

visualize

20

In general, DNA has a _____ appearance and therefore must be ____to be viewed.

transparent, stained

21

DNA can also be separated by size using a technique called

gel electrophoresis.

22

Gel electrophoresis operates on the principle of separation of molecules based on ____ and_____

charge, size

23

When an electrical current is applied in a solution, molecules of solute with a positive charge will migrate to the

cathode

24

cathode= (an electrode through which current flows ___ the electrophoresis chamber, in this case, ____)

FROM, negative

25

anode=(an electrode through which current flows the _____ into the electrophoresis chamber, in this case, _____).

other way, positive

26

molecules of solute with a negative charge will migrate to the

anode

27

In gel electrophoresis, instead of passing an electrical current through a solution, a _____ is used.

gel medium

28

Common gel-forming materials are _____and ______

polyacrylamide, agarose

29

Agarose is a ______ and will be used for this experiment.

polysacchride

30

The agarose gel acts as a type of molecular ____ to facilitate____ of smaller fragments of material.

sieve, separation

31

The gel is placed into a box between the two electrical components:

cathode, anode

32

A small amount of each sample is added to each "____" or _____ at the top of the gel.

well, pre-cast notches

33

Each sample has the DNA sample along with a mix of _____dye or "____ dye" and usually _____

loading, tracking, glycerol

34

The glycerol in the mix weighs the DNA into the well so that it won't rise up and migrate out of the well and the tracking dye allows the researcher to view the progress of the DNA as it ____ through the gel.

moves

35

Molecular weight of DNA proceeds from ____ to ______

heaviest, smallest

36

extract DNA from your cheek cells

____ containing solution is ‘swished’ in the mouth.

salt

37

extract DNA from your cheek cells

The salt dissociates in the water into ____ and ____

Na+, Cl-

38

extract DNA from your cheek cells

The Na+ is attracted to the ______charge of the DNA _________

negative, sugar phosphate backbone

39

extract DNA from your cheek cells

The Na+ is attracted to the negative charge of the DNA sugar phosphate backbone; however this attraction is limited by the presence of _____

water

40

extract DNA from your cheek cells

water is highly ____ and actively_____ to bind with the _____ ions

polar, competes, charged

41

extract DNA from your cheek cells

_____solution containing a detergent and protein degrader are added to dissolve and degrade both ____ and _____

lysis, lipids, proteins

42

extract DNA from your cheek cells

Lysis solution containing a detergent and protein degrader are added to dissolve and degrade both lipids and proteins. This ultimately releases _____into the solution.

DNA

43

extract DNA from your cheek cells

Alcohol ( _____ solution) is added to the mixture.

polar

44

extract DNA from your cheek cells

Alcohol allows for the interaction of the ___ and______charge of the DNA causing a “______” charge of the DNA.

Na+, negative, neutral

45

extract DNA from your cheek cells

The neutral charge makes the DNA ____ and ____ it will precipitate out of the alcohol solution.

nonpolar, subsequently

46

extract DNA from your cheek cells

The neutral charge makes the DNA nonpolar and subsequently it will precipitate out of the alcohol solution. The DNA can then be ___ and______ (dissolved) in water.

collected, re-suspended

47

Restriction enzymes can also give a unique_____ of DNA.

fingerprint

48

Restriction enzymes cut at specific sites within the _____

DNA

49

Restriction enzymes cut at specific sites within the DNA. These sites are usually ____base pairs in length.

4-6

50

Lab staff will prepare DNA digested with the restriction enzymes (2)

EcoRI or Hae III

51

5’-GAATTC-3’

3’-CTTAAG-5’

what restriction enzyme is this

EcoRI

52

5’-GGCC-3’

3’-CCGG-5’

what restriction enzyme is this

HaeIII

53

PCR products are ______pieces of DNA.

amplified

54

DNA consists of a series of nitrogenous base molecules held together by _________ bonds

weak hydrogen

55

if a segment of DNA is diagrammed without the _______and ______

sugars, phosphates

56

look at the linear sequence of bases on each of the strands. Compare the bases in the upper DNA strand to those in the lower strand.

C C A T G G

G G T A C C

Can you discover any relationship between the upper and lower strands?

palindromes

57

When such a sequence is looked at together with its complementary sequence , the group reads the same in both directions. this sequence is called _______ and are fairly common along the DNA molecule

palindromes

58

viruses called _______ are major enemies of bacteria

bacteriophages

59

these viruses (bacteriophages) infect bacteria by _____ their own DNA into bacteria to ___ the bacteria to_____

injecting, force, multiply

60

bacteria have responded by evolving a natural defense called ______ to cut up and destroy the invading DNA

restriction enzymes

61

bacteria prevent digestion of their own DNA by ______ certain DNA bases within the enzyme recognition sequence

modifying

62

bacteria prevent digestion of their own DNA by modifying certain DNA bases within the enzyme recognition sequence, which allows them to _____ their own DNA while _____ foreign DNA

protect, cutting up

63

bacteria prevent digestion of their own DNA by modifying certain DNA bases within the enzyme recognition sequence, which allows them to protect their own DNA while cutting up foreign DNA. this could be considered a very _______________

primitive immune system

64

restriction enzymes search the viral DNA for specific _____ sequences of base pairs such as GAATTC, and_____ the DNA at these sites.

palindromic, cut

65

the actual sequence of DNA is called a

restriction site

66

some restriction enzymes may leave a short length of unpaired nucleotide bases called a "____" end,

sticky

67

some restriction enzymes may leave a short length of unpaired nucleotide bases called a "sticky" end, at the ____ site where they cut

DNA

68

some restriction enzymes may leave a short length of unpaired nucleotide bases called a "sticky" end, at the DNA site where they cut, where as other restriction enzymes make a cut across ____ strands

both

69

some restriction enzymes may leave a short length of unpaired nucleotide bases called a "sticky" end, at the DNA site where they cut, where as other restriction enzymes make a cut across both strands creating ____ stranded DNA fragments with "____" ends

double, blunt

70

the restriction enzyme EcoRI cuts between ___and ____in the palindromic sequence GAATTC

G, A

71

an important feature of restriction enzymes is that each enzyme only recognizes a specific _____ and cuts the DNA only at the specific sequence of _____

palindrome, bases

72

a palindrome sequence can be repeated a number of times on a strand of DNA and the specific restriction enzyme will cut all those palindromes no matter what _____ the DNA comes from

species

73

The linear DNA molecule is represented by ___ line although in actuality DNA has ___strands

1, 2

74

the larger the size of the particles however the ____they are ____ through the gel.

slower, strained

75

after a period of exposure to the electrical current the DNA fragments will ___ themselves out by ____

sort, size

76

fragments that are the ___ size will tend to move together through the gel and form _____

same, bands