Brainscape's Knowledge GenomeTM

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


See full index

Cell and Molecular Biology > Recombinant Technology > Flashcards

Recombinant Technology Flashcards

You may prefer our related Brainscape-certified flashcards:
AP® Biology flashcards Biology 101 flashcards
1
Q

What is the role of restriction enzymes?

A

They cut DNA in to manageable sizes called restriction fragments

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

Restriction enzymes usually:

A

1) Act as dimers and recognise short palindromic DNA sequences
2) Have precise recognition sequences
3) Some leave overhangs but others cut the DNA flush in the middle and are called blunt restriction enzymes

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

How can fragments be separated?

A

Gel electrophoresis

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

Which elctrode is DNA attracted to and why?

A

Positive as it has a negative charge

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

What is ethidium bromide used for?

A

To stain the DNA

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

What is ligase?

A

An enzyme which joins DNA fragments to create recombinant DNA

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

Cohesive termi are also known as what?

A

Sticky ends - this is because they can hybridize

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

How is recombinant DNA made?

A

Ligase is used to connect two DNA fragments

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

What are plasmids?

A

Small, circular, extra chromosomal DNA that occurs naturally in bacteria
Have they’re own origin of replication
Usually carry antibiotic resistance genes

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

How many kilobases does a plasmid hold?

A

30 kilobases

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

What is transformation?

A

Involves mixing bacteria with the plasmid DNA and creating temporary holes in the cell membrane by electroporation

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

What are competent bacteria?

A

Bacteria ready to take up new DNA

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

How are transformed bacteria selected for?

A

By using antiobiotic plates ie testing for antibiotic resistance

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

Where do we get the original DNA?

A

We make a library of genomic clones containing everything

OR a library of cDNA clones

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

What is the advantage of genomic libraries?

A

Contains all the regulatory sequences allowing one to study transcriptional regulation

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

What is the advantage of cDNA libraries?

A

Way to identify which genes are expressed in a disease tissue compared to a normal tissue

17
Q

What is the transcriptome?

A

Genes that are expressed in the original tissue

18
Q

To isolate single clones from the mixed population it is important that there is only:

A

1) One insert into each plasmid
2) One plasmid in each bacteria
3) One bacteria starts with a colony

19
Q

What are ESTs?

A

Expressed sequence tags

20
Q

How are genomic libraries made?

A

You purify and digest chromosomal DNA with restriction enzymes

21
Q

How is sequencing done?

A

1) Denature the template so it is single stranded
2) Cool with the primer
3) Start DNA synthesis reaction with DNA polymerase and dNTPs

22
Q

What is the role of ddNTPS? (dideoxynucleotides)

A

Once they are added DNA polymerase can no longer extend

23
Q

What is automated sequencing?

A

Up to 1000 nucleotides can be read in one reaction

ie ddNTPs are labelled with a fluorescent dye which can be read allowing us to do a single reaction

24
Q

What is done if you want to sequence DNA larger than 1kb?

A

Progressive sequencing

25
Q

What is progressive sequencing?

A

Ends of clones are sequences using primers from the vector
Primers are designed based upon the new sequence and another round of sequencing is performed and so on until the sequences meet in the middle

26
Q

What is shot gun sequencing?

A

Where a genomic plasmid library is made and the ends from each clone is sequenced

27
Q

What are the advantages of shotgun sequencing?

A

Requires no thought and it is automated

28
Q

What are the disadvantages of shot gun sequencing?

A

Need to sequence more than 6 times the genome to get large contigs and there will always still be gaps

29
Q

How many base pairs are in the human genome?

A

3.2^9 base pairs

30
Q

How long did it take to sequence the human genome?

A

13 years (1990 to 2003)

31
Q

How long does it take to sequence a genome now?

A

Around 56 hours

Cell and Molecular Biology (17 decks)

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