Brainscape's Knowledge GenomeTM


Top Flashcards (Certified)
All Flashcards

Biology > Recombinant DNA Technology Word Stimulants > Flashcards

Recombinant DNA Technology Word Stimulants Flashcards

1
Q

Restriction enzymes abbreviated name

A

Restriction endonucleases

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

Restriction enzyme function

A

Catalyse hydrolysis is phosphodiester bonds between sugar-phosphate, double stranded DNA
Cuts at a point in a short DNA sequence (recognition site)

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

Recognition site

A

Specific to each restriction enzyme

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

Sticky Ends

A

Same reverse complements
Form hydrogen bonds with complementary bases to sticky ends of other DNA fragments
Cut by the same restriction enzyme

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

Blunt ended filaments

A

Cut at the same place within recognition site

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

Restriction fragment length polymorphism (RFLP)

A

Lengths of DNA produced from restriction enzyme activity

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

Gel electrophoresis purpose

A

Separate RFLP lengths by size

Gene fragment cut out from gel - available to transfer into organisms DNA

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

mRNA > cDNA

A

mRNA - retrovirus

Reverse transcriptase

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

complementary DNA

A

cDNA
Complementary to parent mRNA
No sticky ends

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

cDNA > double stranded

A

+ nucleotides

+ DNA polymerase (catalysed nucleotide binding with complementary bases)

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

Genetically modified organisms

A

GM
Genes transformed
Genes from host cell > new organism
(Universal genetic code , transcription, translation mechanisms)

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

Vector

A

Bacterial plasmids
Piece of DNA to which gene is inserted
> mixed hybrid molecule (recombinant DNA)

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

Sticky ends in vivo

A

Makes insertion possible

Same restriction enzyme cuts host DNA +Vector DNA (bonds complementary)

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

Gaps in sticky ends

A

Filled by phosphate groups

Catalysed by ligase

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

Desirable > vector steps

A

Restriction enzyme cut DNA containing gene > fragments (sticky ends)
Same restriction enzyme cuts vector DNA
Complementary sticky ends
DNA fragments mixed with cut plasmids + ligase
Complementary sticky ends bond (sealed by ligase)

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

Vectors > host cells

A

Bacterium E. coli + plasmids added to hot liquid
Contains CaCl2
Ca2+ : increase permeability of bacterial plasma membrane to plasmids
1% of bacterial cells take up plasmids

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

Marker genes purpose

A

Identify host cells that carry desirable gene

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

Marker gene Agar

A

Bacteria mixture : Unaltered plasmids / genetically engineered plasmids
Plated on agar
Bacteria without plasmid - do not grow/ form colonies
Unaltered plasmid / GM plasmids grow, form patterns on afar

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

Replica plating

A

Pattern colonies stamped on sterile nylon cloth
Transferred to agar plate
Unaltered plasmids grow / form colonies

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

Identifying plasmids carrying gene

A

Compare agar + replica plate patterns

Colonies Cut + cultured

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

Fluorescent / enzyme markers

A

Is used to identify plasmid carrying gene
Now used
Quicker
More efficient

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

In vivo use

A

Follow in vitro

Testing drug safety

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

In vivo

A

Correspond to real life conditions

24
Q

Disadvantages

A

Complicated
Expensive
Less sensitive

25
In vitro gene cloning purpose
Amplified DNA
26
Polymerase Chain Reaction purpose
PCR Form multiple copies of DNA Cloned Cycle doubles amount of DNA
27
PCR conditions
Dead cells | Lab ware
28
PCR uses
Biological and medical research (evolutionary relationships, diagnosis of diseases / genetic disorders / cancer) Material for analysis (crime scene)
29
PCR step 1
Mixture: primers + double stranded DNA + nucleotides + heat stable DNA polymerase
30
Primer
Short strands of DNA
31
PCR step 2
Heat to 95-98c | Splits double stranded DNA > Single stranded templates (x2)
32
PCR step 3
Cool to 50-65c | Template DNA hydrodizes with complementary primer
33
PCR step 4
Heat mixture to 72 c Addition of complementary nucleotides to template DNA Length of tDAN extends from primers Two copies of double stranded identical DNA
34
In vitro Advantages
More sensitive Simpler Cheaper Variables more easily controlled
35
In vitro disadvantaged
Doesn’t correspond well to real life conditions | May produce mis leading results
36
DNA probes
Single strand of DNA | Base sequence complementary to identifying gene
37
In vitro test steps
Restriction enzyme added to test material DNA cut > single stranded fragments Complementary probe added to fragment mixture Probe labelled with radioactive tag Probe hybridises with cDNA Tag identifies probe/gene hybrid on gel (after gel electrophoresis)
38
DNA probes + DNA hybridisation
Possible to identify + locate specific alleles of genes | Identify genetic disorders (mutant alleles)
39
Genetic screening
Uses DNA sequencing, probes + hybridisation individuals Diagnose individuals : mutant genes/genetic disorders; variants of genes + lifestyle increase susceptibility to particular diseases
40
Informed consent
Advice from expert Informs individual of benefits / limitations of screening Before testing permission
41
Genetic counselling
Basic features of genetic disorder / gene linked disease Probability of developing Probability of child inheritance Options for management, prevention, reduce effects
42
Personalised medicine
Specific medicine based on genetic screening
43
Genetic fingerprint
DNA unique to individual
44
Variable number tandem repeats
``` VNTRs Sequences of non coding DNA repeated base sequences Lengths vary person to person Each person 50-100 types of unique VNTRs - genetic fingerprinting basis ```
45
Genetic fingerprinting use
Criminal investigation - compare DNA from crime scene with suspects Genetic variability in populations Parent of child Medical diagnosis Guide breeding programmes (prevent inbreeding)
46
Genetic fingerprinting steps
``` Extraction Cutting (digestion) Separation Transfer Hybridisation Display Analysis ```
47
Genetic fingerprinting : extraction
DNA extracted from cell samples
48
Genetic fingerprinting : cutting / digestion
Restriction enzymes Cut double stranded fragments Some carry VNTRs
49
Genetic fingerprinting : separation
Helen electrophoresis Pattern of invisible bands DNA broken into single strands - block of gel immersed in alkaline solution
50
Gel electrophoresis equipment
``` -be cathode +be anode Wells of DNA fragments Gel blocks DNA fragment (negative) movement to anode ```
51
Electrophoretogram
Dyed / radioactively labelled | Barcodes
52
Genetic fingerprinting : transfer
Southern blotting | Single stranded DNA fragments transferred onto nylon membrane
53
Genetic fingerprinting : hybridisation
Nylon membrane immersed in DNA probe solution Probe complementary to core nucleotide sequence of VNTRs Probe Labelled with alkaline phosphatase Probe and complementary DNA combine - DNA hybrid
54
Genetic fingerprinting : display
Nylon membrane covered with phosphate containing substrate X Ray film Phosphatase catalyses phosphate removal Substrate fogs X-ray film X-ray shows band pattern - identifies hybrid position
55
Genetic fingerprinting : analysis
Compare bar codes

Biology (25 decks)

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