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Biology Lab > Lab 10 results > Flashcards

Lab 10 results Flashcards

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1
Q

restriction enzymes

A
  • cut only some selective location
  • found in nature (bacteria)
  • they only cut if they see some sequence
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2
Q

palindromic sequence

A

5’ GAATTC 3’
3’ CTTAAG 5’
its the same cause you always read from the 5’ to 3’ end so they are still the same sequence

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3
Q

sticky ends

A

left side- G
CTTAA

right side- AATTC
G

single strands can stick to each other

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4
Q

blunt end

A

no single strand exactment; line cut right down the middle of the sequence

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5
Q

which type of end, sticky or blunt, is better for ligation

A

sticky end

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6
Q

how are restriction enzymes named

A

named after the bacteria they come from
- fourth letter represents strain
- first letter represents genus

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7
Q

what does the buffer contain

A
  • some salt to control ionic strengths
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8
Q

what is the one of the most important salts in the buffer

A

Mg2+
make sures the enzyme is active

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9
Q

2 ul of 10X buffer meaning

A

10 times as concentrated as 1

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10
Q

unit of enzyme

A
  • called a unit
  • usually use 10-20 units
  • don’t measure them by how much
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11
Q

what is an enzyme

A

protein so they can be denatured easily

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12
Q

why do we use 37 degrees C

A
  • human body temp
  • temp most enzymes are active in
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13
Q

what is in the loading dye/buffer

A
  • chemical for controlling pH
  • EDTA to stop DNA from breaking down by getting rid of Mg and Ca
  • glycerol to make sample heavier so it will drop to bottom of well
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14
Q

why do you add the loading dye/buffer

A
  1. so you can see your sample drop in well
  2. dye migrates in same direction as DNA because both are (-)
  3. migrate at certain position depending on DNA
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15
Q

what is the purpose of gel electrophoresis

A

used to analyze DNA

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15
Q

what are some details of agarose

A
  • not digestive
  • when heated in water and then cooled, forms a gel and DNA can migrate through the holes
  • polymer of carbs
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16
Q

what did we use to perform the agarose gel electrophoresis

A

buffer and agarose gel

17
Q

if everything stays the same except agarose, what will occur

A

DNA will migrate to lower % agarose because DNA will move faster

18
Q

if everything stays the same except for use of a different molecule, what will occur

A

small DNA fragment- migrate faster
large DNA fragment- migrate slower

19
Q

if everything stays the same except for voltage, what will occur

A

higher voltage- DNA will migrate faster
too high voltage- melt gel

20
Q

what does a low voltage usually provide

A

a slightly better resolution

21
Q

what are some parameters affecting gel electrophoresis

A
  • agarose %
  • DNA fragment size
  • voltage
22
Q

why do we use SyBr safe

A

need something to stick to DNA to become visible

23
Q

what percentage gel did we make in lab

A

1%
- usually universal concentration
- will analyze from a few 100 base pair to 100 kilo base pairs

24
Q

0.7% gel

A

used for something bigger than 2 kilo base pair

25
Q

2.0% gel

A

used for something smaller than 200 base pair

26
Q

what happens if you make a gel higher than 2%

A

gel will become too sticky

27
Q

what happens if you make a gel with too low of a %

A

gel can brake easily

28
Q

what buffer can be used

A

TAE (EDTA) or TBE

29
Q

TBE

A

used for small DNA

30
Q

TAE

A

more universal

31
Q

what end is DNA attracted to

A

the positive end since it is negative
- opposites attract

32
Q

what does the restriction digestion buffer contain

A

Mg2+

33
Q

when using a 5x stock buffer to make a solution of 20ul, how many ml of stock buffer is needed

A

4

34
Q

if everything else remains the same, DNA migrates faster in agarose gel when

A

the voltage is higher

35
Q

what is the least likely name of restriction enzyme

A

pfu

36
Q

a typical agarose gel is what %

A

1%

37
Q

what is the typical temperature for enzyme digestion in lab

A

37 degrees celcius

38
Q

what does the loading buffer not contain

A

Mg2+

39
Q

how is SyBr safe added

A

directly into the gel

Biology Lab (23 decks)

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