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WACE Human Biology > Biotechnology > Flashcards

Biotechnology Flashcards

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1
Q

What is Gel Electrophoresis?

A

A technique used to seperate DNA fragments according to size, used in DNA profiling to give a unique pattern/sequence

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2
Q

What is Polymerase Chain Reaction?

A

A technique used to amplify small quantities of DNA of a specific DNA region

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3
Q

What is Restriction Enzyme Digestion?

A

The process of cutting DNA at specific sequence / site

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4
Q

What is Profiling?

A

Provides a unique pattern of bands from a DNA sample, used to compare samples

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5
Q

What is DNA Sequencing?

A

DNA sequencing is the process of determining the sequence of nucleotides e.g. AATTT

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6
Q

What is DNA Extraction?

A

Process by which DNA is removed from the nucleus and from the cells

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7
Q

What are Recognition Sites?

A

An enzyme will always cut the DNA at a point where there is a specific sequence of bases

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8
Q

What does DNA ligase do?

A

An enzyme found in E.coli bacteria that is used to ‘glue’ together short strands of DNA during replication

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9
Q

What are Plasmids?

A

Small circular strand of DNA distinct from a cell’s chromosomal DNA, it is able to replicate independently within a cell

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10
Q

What does ‘Ligase’ refer to?

A

An enzyme capable of combining two small components of single strand DNA into one single structure

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11
Q

What are the steps in Polymerase Chain Reaction?

A
  1. DNA is denatured at 94-96 degrees, a double strand is separated into 2 single strands
  2. Annealing primers activate between 50-60 degrees, temperature drops slightly and then primers attach/anneal and bind to each strand and their complementary bases, a new DNA is then synthesised
  3. At 72 degrees Taq polymerase attaches to the primer and extends/syntheses the new DNA strand
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12
Q

What is the application of PCR?

A
  • Detecting hereditary diseases
  • Detection of viral diseases
  • Forensic science - when needing to amplify small amounts of blood, semen, hair etc
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13
Q

Explain the process of Gel Electrophoresis

A
  1. Make the agarose gel
  2. Add comb to the gel and allow it to set
  3. Add dye to the DNA sample
  4. The buffer (liquid) is added to cover the gel
  5. DNA sample is loaded into the wells of the gel
  6. An electric current is applied
  7. DNA is negatively charged and therefore the DNA fragments move from the negative end to the positive end as the DNA is attracted to the positive end
  8. Smaller DNA segments move faster and farther than larger DNA fragments
  9. Gel is stained with DNA binding dye for a few minutes
  10. Gel is placed under UV light and the sequence will fluoresce, showing the band sequence
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14
Q

Explain the process of DNA sequencing?

A
  1. The mixture is first heated to denature the template DNA (separate the strands)
  2. The temperature is then lowered slightly so that the primer can attach/enter
  3. Temperature is then slightly raised and the enzyme binds to the DNA and elongation occurs
  4. DNA will elongate until a dideoxynucelotide binds then the sequence wills top as no more can bind as the dideoxynucelotide lacks OH, this results in the collection of DNA strands in different sizes and lengths
  5. The DNA then goes through a polyacymide gel which s more porous in a DNA sequencer
  6. The DNA sequencer then analyses the gel and the fragments migrate according to size and each is detected as it passes through a laser beam light, each dideoxynucelotide emits a different colour and is recorded as a colour band on a stimulated gel image which gives us the exact DNA sequence
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15
Q

What is Gene Therapy?

A
  • Aim to treat or cure genetic abnormalities by replacing faulty genes with healthy ones
  • The genes can be used as the treatment
  • Gene therapy is taking place with single gene disorders such as cystic fibrous and Huntington’s disease
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16
Q

What are Gene Probes?

A
  • A single stranded DNA labelled with a radioactive or fluorescent marker to detect the presence of specific DNA sequences on another DNA or RNA molecule
  • Can be used for detection of hereditary diseases. the probe is made to specifically bind to the specific DNA sequence of the disease
17
Q

What is Restriction Enzyme Digestion?

A
  • Cuts the DNA at a specific sequence / site
  • Some produce a straight cut at the sequence, this is when a restriction enzyme makes a clean break across the two strands of DNA to produce a blunt end which is when both strands terminate in a base pair
  • Some produce a staggered cut resulting in fragments with sticky ends, a sticky end is a stretch of unpaired nucleotides in the DNA molecule that overhang at the break in the strands
18
Q

What is Tissue Engineering?

A

Tissue Engineering refers to restoring healthy tissues or organs and eliminates the need for organ/tissue transplants or artificial transplants, it requires abundant cell supply and can be induced to grow on a scaffold of natural or synthetic material to produce 3D tissue

19
Q

What is Recombinant DNA?

A
  • Used in Genetic Engineering, requires a vector
  • It involves isolating the DNA and amplifying it and insertion of gene/DNA segment into transgenic bacteria (foreign to the organism or may be modified)
  • Can be used to replace genes e.g. diagnosis and treatment of genetic disorders, manufacture large quantitates of pure proteins such as insulin, human growth hormone, factor VIII
20
Q

What are the Steps in Recombinant DNA?

A
  1. Isolate the gene of interest and cut out using a restriction enzyme
  2. Isolate a plasmid from bacterial cell and cut it with the same type of restriction enzyme used in step 1
  3. Splice the human DNA into the plasmid using DNA ligase enzymes to join the sticky ends
  4. Treat the bacterium so it takes up the recombinant plasmid and once this is successful, the bacterium will multiply so that either the human gene or product of the gene can be used
21
Q

How is Haemophilia an example of Recombinant DNA?

A
  • Haemophilia is a genetic disorder in which the blood clotting protein factor VIII is affected
  • VIII is in small supply and therefore a person suffering from haemophilia is unable to form blood clots accurately and there is a risk of death from excessive bleeding
  • In the past factor VIII was obtained by blood donors however there was a risk of contamination transmission of viral diseases such as HIV and Hepatitis C
  • Recombinant DNA techniques overcame this issue and therefore is a massive advantage
  • It is cultured in mammalian cells and is considered highly effective in the control of excess bleeding

WACE Human Biology (15 decks)

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