Topic 6 Flashcards
(123 cards)
What does DNA profiling rely on
The fact that everyone’s DNA bar twins is unique
What are introns
They are non-coding blocks and are inherited in the same way as genes
What do introns contain
They contain STR satellites that are repeated several times
Although the same str occur at the same loci on both chromosomes of a homologous pair how do they differ
They differ in the number of times they are repeated
Why is the variation in the number of repeats of str on a loci good
Scientists use this to create unique DNA profiles
Gives the steps in obtaining a Tissue sample
A sample of DNA is collected from biological tissue i.e. Blood smear
The tissue sample is broken using a buffet solution that contains a salt and detergent the cell memebranes are disrupted
The DNA is separated from the rear of the cell debris by flirtation and centrifuging
Proteins are removed due the fact that protease enzymes are incubated and cold ethanol is used to precipitate the DNA
What two ways can be used in creating DNA fragments
Restriction enzymes or Polymerase chain reaction
Where do restriction enzymes come from
They are found in bacteria and its name is reflected by the bacteria it comes from
What do restriction enzymes do
They cut DNA at specific recognition sites at either side of the DNA which means the fragment is intact but cut from the rest of the genome
When do restriction enzymes cut a DNA sample
When their specific restriction sequence occurs
What are DNA primers
They are DNA sequences that contain fluorescent tags and are complementary to the DNA adjacent to the str
Describe the steps of PCR
A the DNA from a sample along with DNA polymerase, DNA primers with markers and nucleotides are added to a reaction tube
At 95 degrees the DNA separates into two strands
At 55 the DNA primers attach to the start of the str repeated sequence on each strand
At 70 DNA polymerase attach the two nucleotides together which extends the extend the DNA from the primer and the str repeated sequence along with the DNA adjacent are replicated DNA polymerase is attached
This cycle repeats its self producing a large number of DNA fragments
What method is used to separate the DNA fragments
Gel electrophoresis or southern blotting
What technique is used to separate DNA fragments
Gel electrophoresis Its done accordance to size
Describe what occurs in gel electrophoresis
The DNA fragments are placed in a gel of garose or polyacrylamide
The gel has electrodes connected to it and it produces a Pd across it and it’s placed in a buffer solution
The DNA fragments are negative charged and move through the gel in accordance to size and charge, so smaller sequences travel to the positive electrode faster
Fragments are measured in base pairs
What technique is used to visualise fragments
Southern blotting or
Gel electrophoresis
What is southern blotting used for
To transfer DNA fragments to a more resilient nylon membrane as gel is to fragile
Describe stages in southern blotting and when it’s used
After restriction enzymes
The membrane and dry absorber paper are place on the gel directly to draw the buffer solution up alive with DNA fragments denatures strands from double to single
- The membrane is incubated with excess of labelled DNA probe (either radioactive or fluroescent
- to visualise with a radioactive probe, the membrane is dried first and placed to an X ray film,if the probe has bound with DNA, the film go black
If the probe is fluorescent,the postion on the membrane is visualised under uv light
In southern blotting what is the DNA probe and what does it do
It’s a short DNA section complementary to the base sequence of target DNA and they are either radioactive or labelled with fluorescent marker
In a DNA profile why does a single bond occur
When a persons maternal and paternal chromosome have the same number of stars in a certain locus
When do 2 bands occur on a DNA profile
If the two chromsomes have a different number of repeats at a locus
In visualising DNA when do we use gel electroresis
After PCR
Describe the stages in Gel electrophoresis in analysing DNA fragments
The DNA primers have fluorescent tags
The DNA fragments with their tags pass through a laser in the gel,this causes the the dye in the tags to fluoresce and the coloured light is detected
This gives a time taken fragments to pass through the gel and the time length can be calibrated with the fragment size
Str loci can be analysed by using the tags that fluroresce at different wavelengths as they give different colours for each str loci
A computer processes this info from the sector and displays the results of this on s graph
What determines the size of the fragment t
Number of base pairs