Manipulating genomes Flashcards
what is PCR used for
to select a fragment of DNA and amplify to produce millions of copies
describe the polymerase chain reaction
step 1: create a mixture of free nucleotides, primers and DNA polymerase
step2: DNA mixture is heated to 95oc to break hydrogen bonds between bases, the mixture is then cooled to 54oc so that primers can anneal to the strands
step 3: reaction is heated to 72 so DNA polymerase can function and complementary base pairing can occur
step 4: two new copies of the fragment of DNA are formed and the cycle restarts
what are palindromic sequences
antiparallel base pairs (base pairs that read the same inapposite directions)
what do restriction enzymes do
recognise specific palindromic sequences (known as recognition sequences) and cut (digest) the DNA at these places)
how is DNA cut
through hydrolysis reaction
Sometimes a cut leaves sticky ends what does this mean
they are small tails of unpaired bases at each end of the fragment which can then be used to anneal the DNA to an other fragment with complementary base pairs
Describe the process of electrophoresis
- agarose gel is poured an left to solidify
- row of wells is created closest to the negative electrode (cathode)
- a buffer solution is then added
- using a micropipette add same volume of DNA into each well
- add a power supply to create electrical current
- DNA fragments will move to the positive electrode with smaller pieces moving further and faster
- remove the gel tray and stain the DNA fragments to make them visible
what is electrophoresis
procedure that uses an electrical current to separate out DNA fragments based on size
what is DNA profiling used for
forensics and medical diagnosis
what is DNA profiling
the number of times a sequence us repeated at different specific places in a persons genome is analysed using electrophoresis
what is genetic engineering
the manipulation of an organisms DNA
what is recombinant DNA
DNA formed by joining together DNA from different sources
how is genetic engineering carried out
- extract a DNA fragment that contains the desired gene, using restriction enzymes
- Insert the DNA fragment into vector DNA
- isolate the vector DNA
- vector DNA is cut open using restriction enzymes using the same one that was used to isolate the DNA fragment so that the sticky ends are now complementary
- the vector DNA and DNA fragment are mixed together with DNA ligase which joins the sugar phosphate backbones of the two bits of DNA
- recombinant DNA is formed
- the vector with the recombinant DNA is used to transfer the gene into the bacterial cells
what is an example of genetically modified plants
soybean plants have been genetically modified to be insect resistant
what are the positive and ethical issues concerning GM soybean plants
+ reduce the amount of chemical pesticides used, can be more nutritious
- may encourage monoculture which decreases biodiversity and makes them vulnerable to disease
- could interbreed with wild plants creating weeds resistant to herbicides
-expensive for poorer farmers