Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Biol 207 > Lecture 8 > Flashcards

Lecture 8 Flashcards

1
Q

What does Recombinant DNA technology allow for?

A

Allows for scientists to identify, isolate and combine DNA fragments from any organism to obtain the desired properties in new DNA molecule that can be maintained in bacterial

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

What does Polymerase chain reaction (PCR) allow for?

A

Allows for the amplification of small to medium-sized DNA fragments from minute amounts. RNA can be amplified after copying it into DNA

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

What does DNA sequencing allow for?

A

Allows the determination of the nucleotide sequence of a DNA fragment (and RNA after copying it into DNA).

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

What does CRISPR/Cas genome editing all for?

A

Allows for the precise manipulation of DNA and in the case of Cas13, RNA, in a wide range of organisms. The Cas9 enzyme cuts DNA in a precise location because it is guided to the the target sequence through the presence of a “single guide RNA”

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

What can CRISPR be used to generate?

A

Simple mutations or complex gene editing (gene replacement) approaches

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

What is a mutation?

A

Change the DNA sequence of a given gene to create a mutant organisms. This includes classic (random) and modern (targeted) mutations

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What is Transgenesis?

A

Introduce a new piece of DNA to the genome to create a transgenic organism

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

What does Transgenesis result in?

A

Because a new piece of DNA is introduced this causes the organism to produce a new type of protein that has some new property

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

What is Gene editing?

A

A fairly new technique (CRISPR/CAS9) is a blend of mutations and transgenesis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What are the 2 ways you can clone a gene?

A

Using PCR or retrieve it from a DNA library

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

What can DNA Libraries be used for?

A

Genomic DNA or cDNAs

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

What is the idea behind DNA libraries?

A

The idea is to have an entire genome or transcriptome represented in individual plasmid DNAs (vector DNAs)

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

How are different fragments of DNA created in DNA Libraries?

A

Genomic DNA is digested by restriction enzymes to create different fragments

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

How do Restriction enzymes know where to cut?

A

They recognize specific DNA sequences

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

What occurs in Reverse Transcriptase?

A

An RNA template is used to make DNA

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

Who invented PCR?

A

Mullis

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

Why are Thermocyclers necessary in PCR?

A

It is needed to tear the double stranded DNA apart

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

How do we clone our GOI into a suitable vector?

A

Restriction enzymes recognize specific DNA sequences and cuts it to create sticky ends (ends with overhangs). Then the vector DNA and PCR fragment are cut to make the DNA ends compatible and ligase is added to fuse them together

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

What are sticky ends?

A

When DNA is cut by restriction enzymes they create 5’ overhangs called sticky ends

20
Q

When is DNA said to be clones?

A

Upon successful transformation of the recombinant DNA into the bacteria

21
Q

What does Transposase do?

A

Integrates the transposon DNA into new integration sites

22
Q

How is Transposase created?

A

A helper plasmid encodes a function copy of Transposase

23
Q

What does the P-Element contain?

A

The vector with the recombinant DNA that contains recognition sites for the Transposase

24
Q

How is Recombinant DNA added to a fly genome?

A

Both the P-element vector and the helper plasmid are co-injected into the fly embryo which causes insertion of the P-element into the host genome

25
Q

How can we know if integration of a gene into a drosophila is successful?

A

Successful integration can be monitored by expression of the w+ gene (makes red eyes) the hose genome is w- so it has white eyes

26
Q

How does Sanger Sequencing work?

A

A DNA template and normal nucleotides (dNTPs) undergo synthesis by DNA polymerase. In each tube there is a certain nucleotide type of (ddNTPs) in small amounts that will block the activity of DNA polymerase creating different fragments

27
Q

What are the normal nucleotides in Sanger sequencing?

A

dNTPs

28
Q

What are the abnormal nucleotides in Sanger Sequencing?

A

ddNTPs that are missing both OH groups

29
Q

What occurs after replication in Sanger Sequencing?

A

Since the fragments also contain radioactively labelled dNTP we can see their length. The fragments then become denatured and the single strands are separated by gel-electrophoresis

30
Q

What is different about Automated Sanger Sequencing?

A

The ddNTPs are labelled with fluorescent dye and after replication and electrophoresis the fluorescent dyes are detected using a detector. This all takes place in a single reaction tube rather than 4.

31
Q

What does CRISPR stand for?

A

Clustered Regularly Interspaced Short Palindromic Repeats

32
Q

Where was CRISPR discovered?

A

In Bacteria and Archaebacteria

33
Q

What does CRISPR do in Bacteria and Archaebacteria?

A

Serves as memory against phages

34
Q

How does CRISPR help Bacteria and Archaebacteria?

A

DNA from an invading phage is stored in a specific locus if the bacteria chromosome. Should another infection occur from a phage with the same DNA the stored CRISPR DNA is used to identify the invading DNA and cut it via Cas nucleases

35
Q

What are Cas nucleases?

A

DNA degrading enzymes that are guided to their target by CRISPR-derived RNA

36
Q

Why are Cas nucleases known as engineered nucleases?

A

Because the enzyme is recruited to specific locus

37
Q

What would the Cas nuclease result in?

A

A double strand break

38
Q

What is the simplest way that CRISPR allows the double stranded break to be repaired?

A

Non-homologous end joining

39
Q

What will NHEJ cause?

A

Small deletions in the targeted region resulting in gene mutation

40
Q

Why is NHEJ not gene editing?

A

Because we don’t technically have control on what the final sequence will be

41
Q

What did Homologous recombination depend on?

A

The sister chromatid to fix the double stranded break

42
Q

What is it called when CRISPR depends on homologous recombination?

A

Homology-Directed Repair (HDR)

43
Q

What occurs in advanced CRISPR?

A

a donor DNA that is homologous but not identical to the region with the double stranded break is injected. The fools the HR system to use the injected donor

44
Q

What is Cas9?

A

An RNA-binding nuclease

45
Q

What is the single RNA that is incorporated into the Cas9 enzymes function?

A

It serves as a guide to direct the CAS9/RNA complex to DNA that is complimentary to the RNA so that it can introduce a double stranded break

46
Q

Why is the RNA in the Cas9 enzyme called the single guide RNA?

A

Because it guides the complex to the complementary DNA

47
Q

What is the PAM sequences?

A

The genetic sequence adjacent to the RNA-DNA match that needs to present next to the target sequence for initial binding of the Cas9 effector complex

Biol 207 (14 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions