Brainscape's Knowledge GenomeTM


Top Flashcards (Certified)
All Flashcards

BIO 111 > Practial 2 > Flashcards

Practial 2 Flashcards

(72 cards)

Start Studying
1
Q

What are the 3 basic steps to DNA Isolation?

A

1.Cell Lysis
2.Protein Precipitation
3.DNA Precipitation/Cleaning

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Is there a strict protocol for DNA Isolation

A

No, many different protocols may apply, all depends on the tissue, organism, and purpose

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

2 types of DNA

A

Plastid-DNA in mitochondria and chloroplasts
Nuclear- DNA from cells nucleus

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What happens during Cell Lysis

A

We get rid of membranes with a lysis buffer
Note: still a lot of macromolecules and proteins to get rid off*

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What happens during Protein Precipitation

A

The removal of histones and other proteins

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What ways to do protein precipitation exist

A
  1. Protease- which is an enzyme that will degrade proteins
  2. Protein precipitate solution- you add it to the cell and it helps precipitate clumps of all those proteins together
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Why do histones have high priority?

A

Because they have a positive charge and will cancel out DNA which has a negative charge

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What happens during DNA precipitation?

A

We get DNA to precipitate out of solution

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What is done after DNA is isolated in DNA isolation?

A
  1. We test for yield/product
  2. Amplify a particular sequence to see if we were successful in our isolation. If there is lots of DNA, you may see a stringy, white precipitate
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What are enzymes?

A

an organic catalyst/protein that speeds up metabolic reactions

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What is energy of activation? (Eact)

A

-the initial energy investment required to have a reaction start/proceed

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

How do enzymes work?

A

By lowering energy of activation (Eact) levels. They make it so the reactants use/have to obtain less energy and therefore get products faster.

Without enzymes, there is no such thing as life

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Catalized reaction

A

Reaction with enzymes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Uncatalized reaction

A

Reaction without enzymes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Substrate

A

The substance that an enzyme acts on/ fits in the enzyme

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Active Site

A

A shallow depression or groove on an enzyme where the substrate fits

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

How can I tell the enzyme is working?

A

-check for production of products

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

What are some factors that affect enzyme activity?

A

1.Time
2.Enzyme concentration
3.Temp
4.pH

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

how does time affect enzyme activity?

A

more time = enzyme activity increases

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

how does enzyme concentration affect enzyme activity?

A

if it increases, enzyme activity increases

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

how does temperature affect enzyme activity?

A

boiling=denatured, the lower the temp, enzyme activity increases

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

how does pH affect enzyme activity?

A

if pH is outside neutrality, enzyme is denatured. Must remain neutral

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

Is it possible for an enzyme to return to its natural state after being denatured?

A

Yes, occasionally whenever you remove the denaturing agent its possible

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

what happened when the enzyme/potato reacted with H2O2 in an environment w ph of 7

A

lots of bubbles appeared, indicating enzyme is working

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
what happened when the enzyme/potato reacted with H2O2 in an environment w ph of 2
no bubbles, indicates no product
26
what happened when the enzyme/potato reacted with H2O2 in an environment w ph of 12
no bubbles, indicates no product
27
What are the phases of interphase?
-G1 -S -G2
28
What takes place in the G1 phase of interphase?
Increase in ribosomes and mitochondria as well as an increase in the size of the cell
29
What takes place in the S phase of interphase?
DNA replication and histones replicate; histones use the ribosomes/proteins and the DNA replication makes use of the atp from mitochondria
30
What takes place in the G2 phase of interphase?
-Further increases in size -Final Preparations for division -produces 2 copies of DNA
31
Can interphase be seen under a microscope?
No
32
What takes place during prophase?
The chromosomes condense and the nuclear membrane starts to break down
33
What takes place in metaphase?
Sister chromatids line up along equator
34
What takes place in anaphase?
Cohesions are cleared/seperated by enzyme called separase Sister chromatids separate and move along the kinetochore microtubules towards opposite ends of the cell.
35
What takes place in telophase?
It’s the opposite of prophase. Chromosomes begin to unwind and nuclear membrane reforms. We are left with 2 identical sets of chromosomes and 2 nuclei began to form into the cells nucleus
36
Can the cell cycle be complete without cytokinesis?
Yes, it will form a cell with 2 nucleus
37
What takes place in cytokinesis?
Depends on the type of cell; animal or plant, but basically finally separate into 2 individual daughter cells
38
How is cytokinesis accomplished in an animal cell?
The formation of a cleavage furrow which contracts/ tightens the microfilaments of the cell until it is pulled apart
39
How is cytokinesis accomplished in a plant cell?
Vesticles containing cell wall material transport the material to the equator of the cell and start to a cell wall in the middle of the cell until completely and form 2 daughter cells
40
Can prophase be seen in a microscope?
Yes, with the use of a stain. The stain stains for nuclear proteins, look for were the stain isn’t evenly distributed throughout the nucleus
41
What is PCR
A way to amplify a targeted portion of DNA. Involves using varying temperatures
42
How is PCR done?
Used to be done by hand, but now we use a machine called a Thermal Cycler
43
Steps of PCR
1. Denaturing 2. Annealing 3. Extension
44
What is an amplicon?
The products of PCR
45
When A is on one strand,…
It will form 2 H bonds with a T on the other strand
46
When G is on one strand,…
It will form 3 H bonds with a C on the other
47
In what case would we substitute T for U
When talking about RNA
48
What is done during the denaturing step of PCR
The temp is raised to around 95°C, this separates the 2 strands of our template DNA
49
What is done during the annealing step of PCR?
The temp is lowered to around 55°C, this allows your forward and backward primer to bind
50
What is done during the extension step of PCR?
The temp is raised to around 72°C, this allows Taq polymerase to start where the primers are. It will read one strand and form a new complementary strand based of CBP
51
How long do the steps of PCR run?
A specific period of time, anywhere from 30 secs to 2 mins
52
Can the temperatures used during PCR vary?
Yes, depending go the number of factors
53
How long is the initial denaturing of PCR?
2-10mins
54
How many times are the basic steps of PCR done?
30-35x
55
What is the final extension done for?
To allow enough time for TAC polymerase to bind to amplicons and finish any of the amplicons that haven’t been able to
56
At what temperature does the thermal cycler keep the template DNA at once PCR is done?
4°C
57
What materials are needed to perform PCR?
-Template DNA -Primers -Taq polymerase -dNTPs
58
What two materials make the “master mix”
Taq polymerase and dNTPs
59
What is the primers job in PCR?
It targets the segment and dictates what specific sequence your amplicon is going to be/replicate
60
What is the dNTPs’ job in PCR?
A nucleotide that has 3 phosphates used to form master mix
61
Why is the “master mix” used?
It is ideal bc it reduces pipetting and risk of contamination, is convenient, saves time and preempts possible errors in mixin
62
What is Taq polymerase and what is its job in PCR?
It is bacteria taken from bacteria found in high temperatures and it is an ingredient for “master mix”
63
What is Gel Electrophoresis?
A way to separate nucleic acids or proteins, with a focus on DNA only.
64
How does Gel electrophoresis work?
Subjects DNA to an electric field, causing it to migrate towards positive end and separates/filters DNA fragments as it migrated through gel
65
Which fragments pass through the gel quicker; small or large?
Small fragments pass quicker through gel
66
Overview of steps of Gel Electrophoresis
1.Have to make a gel (Casting a gel) 2.Assemble parts and pipette DNA into wells 3.Run gel 4. Resolve any imaging issues
67
Is there a set type of compound and composition of compound that is used to make gel for gel electrophoresis?
No, many different compounds and composition of said compounds can be used
68
What compound is commonly used during gel electrophoresis?
Agarose
69
How would different concentrations of compounds affect the gel?
It can affect the type of separation we see and vary based on pore size
70
How is DNA precipitated from cell in DNA precipitation?
Since DNA is polar because of it’s “phosphate backbone”, we can use ethanol, which is less polar, to help precipitate the DNA out the cell
71
What do enzymes do?
they are involved in bond breaking and/or bond formation
72
How do the kinetochore microtubules shorten?
They depolymerize at the end at which they’ve grabbed the each sisterchromatid and pull them apart

BIO 111 (5 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2025 Bold Learning Solutions. Terms and Conditions