Micro satellite DNA
Generally less than 4 bp
Key Terms
Micro satellite DNA
Generally less than 4 bp
DNA profiling procedure
Extraction
Restriction digestion
Separation of the DNA fragments
Southern blotting
Hybridisation
Seeing the evidence<...
Extraction in DNA profiling
DNA must be extracted from a biological sample and then amplified to develop a profile
How to extract DNA
Add detergent
Will break up csm and nuclear membrane
Add salt to form a ppt
Restriction digestion
Extracted DNA is cut by restriction enzymes to produce restriction frgaments
Use the same no. as VNTR's youre looking for
Separation of DNA fragments
Cut fragments need to separated using gel electrophoresis to produce a banding pattern
Alkali solution is poured over the strands and gel to ...
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| Term | Definition |
|---|---|
Micro satellite DNA | Generally less than 4 bp |
DNA profiling procedure | Extraction Restriction digestion Separation of the DNA fragments Southern blotting Hybridisation Seeing the evidence |
Extraction in DNA profiling | DNA must be extracted from a biological sample and then amplified to develop a profile |
How to extract DNA | Add detergent Will break up csm and nuclear membrane Add salt to form a ppt |
Restriction digestion | Extracted DNA is cut by restriction enzymes to produce restriction frgaments Use the same no. as VNTR's youre looking for |
Separation of DNA fragments | Cut fragments need to separated using gel electrophoresis to produce a banding pattern Alkali solution is poured over the strands and gel to separate them into single-stranded molecules |
Southern blotting | DNA (-ve) from gel electrophoresis is transfereed to a +vely charged membrane e.g. nylon Fragments are irreversibly bound to the blot, whilst maintaining their relative positions on the gel |
Hybridisation and seeing the evidence | DNA probe binds onto the blot at a position where the appropriate DNA sequence is found You can detect the position using autoradiography or use fluorescently marked probes that can be viewed w/ UV light |
DNA probes | Single stranded short piece of DNA with a known complementary sequence to the VNTR Synthesised chemically and is radio-labelled |
Radio labelling | Incorporating a small number of radioactive bases into DNA (nitrogen-15) |
Physical effects of Huntington's disease | Shaking of the hands Awkward gait Loss of muscle control and mental function |
Cause of Huntington's disease | Trinucleotide repeat expansion (CAG) on chromosome 4 35+ repeats = Huntingtons disease mHTT gene is dominant |
What does mHTT do | Death of cells of the cerebrum and cerebellum | Results in atrophy of brain matter |
DNA sequencing | Process of working out the order of nucleotide bases in strand of DNA |
Sanger sequencing | DNA sequencing based on the selective incorporation of chain terminating dideoxynucleotides |
Dideoxynucleotides | Chain terminators inhibitors of DNA Polymerase (lacks -OH on C3) |
High throughput sequencing | New methods of sequencing DNA that are automated, very rapid and cheaper than orig. methods |
Capillary gel electrophoresis | Separates macromolecules such as nucleic acids through capillary action in a capillary tube |
Ingredients for Sanger sequencing | DNA polymerase Primer Free nucleotides Template DNA Dideoxynucleotides (Could be added separately or altogether ) |
Method of Sanger sequencing | Add DNA sample to a tube w/ primer, DNA polymerase and DNA nucleotides and dye labeled ddnucleotides in much smaller amounts Follow steps of PCR (heating, cooling, heating) until a ddnucleotide is added Repeat cycle several times until you can be sure a ddnucleotide has been added to every position of the target DNA Carry out capillary gel electrophoresis Smallest fragment will cross the 'finish line' first then the next. The colours of dyes will be registered one after another on the detector and each colour corresponds to a known base |