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Biology > Manipulating genomes > Flashcards

Manipulating genomes Flashcards

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AP® Biology flashcards Biology 101 flashcards
1
Q

PCR

A

polymerase chain reaction
Amplifies short lengths of dna into millions of copies

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2
Q

Step 1 PCR

A

DNA fragments, primers and free nucleotides join, DNA polymerase
Added to thermocycler
94-96 degrees Celsius causes DNA strands to separate (denature) hydrogen bonds

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3
Q

Step 2 PCR

A

primers join to their complementary bases (55-68 degrees Celsius)
Provides a starting point for DNA polymerase

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4
Q

Step 3 PCR

A

temperature increased back up to 72 degrees Celsius (optimum temp for DNA polymerase)
cycle restarts & DNA amplified
Double each time

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5
Q

Applications of PCR

A

-detecting mutations
-research
-forensic science
-identify viral infections
-monitoring spread of infectious disease
-detection of oncongenes

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6
Q

Method to get DNA fragment

A

some sections of DNA are palindromic
-restriction enzymes recognise specific palindromic sequences
-they cut the DNA at these places because shape of sequence is complimentary to active site

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7
Q

Sticky ends

A

small tails of unpaired bases at each end of fragment
Used to join DNA fragment to another piece of DNA that has sticky ends with complementary bases

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8
Q

Gel electrophoresis steps

A

Extraction of DNA from sample
& fragments made by restriction enzyme
Separation using electrophoresis (small fragments travel furthest)
These bands are colourless but are revealed using radioactive or fluorescent probes
(METHOD TO SEPARATE DNA)

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9
Q

DNA ladder

A

Tool used to compare base length sequences against a known sequence length
Estimation of size

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10
Q

SDS

A

charge detergent used to equalise surface charge
So that it moves on molecular mass not charge

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11
Q

Uses of DNA profiling

A

Aid diagnosis of sickle cell
Guilty/non guilty if a crime
Find father of child
embryo screening

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12
Q

DNA probes

A

small fragment of nucleic acid that is labelled with an enzyme, radioactive tag or fluorescent tag
probe binds to complementary bases
-locates specific gene for genetic engineering
-identify same gene in a variety of different genomes from different species

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13
Q

genetic counselling

A

family history of genetic disease

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14
Q

Genetic engineering

A

genes inserted from one organism into another

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15
Q

recombinant DNA

A

a composite DNA molecule created in vitro joining foreign DNA

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16
Q

GE step 1

A

reverse transcriptase will form single stranded complementary DNA (cDNA)
addition of primers & DNA polymerase
makes cDNA into double stranded DNA
whose base sequences code for original desired protein

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17
Q

GE step 2

A

Plasmids obtained from organisms such as bacteria & mixed with restriction enzymes cut at specific sites
-the cut plasmid has exposed sticky ends
DNA ligase will anneal the free nucleotides to sticky ends

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18
Q

GE step 3

A

Heat shock treatment
-if bacteria are subjected to periods of hot and cold in presence of calcium chloride
-membranes become more porous and allows in recombinant DNA

19
Q

Electroporation

A

high voltage pulse is applied to cell to disrupt the membrane

20
Q

Electrofusion

A

electrical fields help introduce DNA into cells
heals with DNA inside cell

21
Q

Agrobacterium Tumefaciens

A

infects plants

22
Q

DNA profiling

A

procedure
-extract DNA from sample
-Digest sample ( DNA cut into fragments) by restriction enzymes
-separate DNA fragments using electrophoresis
-Bonding pattern produced
patterns compared

23
Q

tandem repeat sequences

A

-non coding regions
-10-100 base pairs long
-all feature same core sequence
-introns/ telemeres satelite DNA
-similar tandem repeat sequences in relatives

24
Q

DNA sequencing

A

Identifying the base sequence of a DNA fragment

25
Q

Fred Sangers

A

use a single strand of DNA as template for 4 experiments in separate dishes
Each dish contains solution made up of 4 bases (ATGC) , DNA polymerase
Each dish has a modified version of one of the DNA bases
(modification = being labelled with a radioactive isotope)

26
Q

terminators

A

the modified nucleotides

27
Q

mixture in reaction

A

-The DNA being sequenced
-mixture of (normal) nucleotides
-one type of terminator nucleotide (modified)
- a primer
-DNA polymerase

28
Q

Genome sequencing

A

-genome cut into smaller fragments (restriction enzymes)
-fragments inserted into BACs
-BACs inserted into bacteria
-Bacteria then divide
(creating colonies of cloned cell)
complete genomic DNA library
-DNA then extracted from each colony & cut up using RE
DNA fragments then put back in order by computers

29
Q

Automated sequencing

A

-Nucleotides are fluorescently labelled with dyes
-Everything occurs in single tube
separation can occur in one lane during gel electrophoresis

30
Q

Pyrosequencing

A

Involves synthesising ssDNA
strand sequenced one base at a time and using light emission which base was added at each time

31
Q

Pyrosequencing steps

A

long length of DNA to be sequenced is mechanically cut into fragments of 300-800bp using nebuliser
- these lengths are often degraded into single stranded DNA (ssDNA)
-these are template DNAs and are immobilised
-Sequencing primer added & DNA is incubated with the (DNA polymerase, ATP sulfurylase, luciferase, apyrase
& adenosine 5 phosphosufate

32
Q

Activated nucleotide

A

(nucleotide with 2 extra phosphoryl groups)
eg TTP incorporated into complementary strand of DNA by using strand as template
As this happens the 2 extra phosphoryls are released pyrophosphate

33
Q

ATP sulfurylase

A

in presence of APS
converts pyrophosphate –> ATP

34
Q

luciferase

A

converts luciferin –> oxyluciferin
generates visible light

35
Q

Bioinformatics

A

study of sequences

36
Q

Benefits of comparing genomes

A

Allow us to compare between species to see evolutionary relationships
Comparing between individuals allows us to tailor medical treatments

37
Q

Process of inserting DNA fragment into a vector

A

Plasmid used as vector & is cut using the same restriction enzymes so that ends are complimentary
-DNA ligase joins fragment and plasmid together

38
Q

Process of inserting vector into host cell

A

host cells mixed with vectors in ice cold solution then shocked to increase permeability of cell membrane (electroporation) which encourages the cells to take up the vector

39
Q

disadvantages of genetic engineering

A

GE seeds would be harder to aquire for poor farmers
GM crops may cause antibiotic resistance

40
Q

advantages of genetic engineering

A

GM animals used to make pharmaceuticals
GM foods have longer shelf life
GM food can have better nutrient content

41
Q

DNA sequencing

A

Knowing the sequence of a gene allow us to predict the sequence of amino acid that makes up the polypeptide

42
Q

applications of gene sequencing

A

comparison between species
evolutionary relationships
variation between individuals

43
Q

synthetic biology

A

science concerned with designing and building useful biological devices and systems.

Biology (22 decks)

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