Summary: Molecular Biology Labo
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1 DNA isolation and start-up PCR
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What is the aim of this experiment?
In thisexperiment we are going toisolate DNA from the cheek using salt wash. Then we will try todetect aDNA polymorphism usingPCR . You willdetect anAlu insertion polymorphism situated onchromosome 8, in theneighbourhood of theTPA gene -
In which genotypes does the PV92 system result?
In three types (++, +-, --) -
How can we seperated the + and - allels?
By size using gel electrophoresis -
How are Alu elements classified?
As SINEs: Short INtersperesed Elements -
What is the lenght of Alus?
Approximately 300 bp -
How many Alu copies does the human chromosomes contain?
About 1 000 000 000 which equal 10% of the total genome. -
Alu is an expample of a so-called "juming gene" what is that?
A transponsable DNA sequence that "reproduces" by copying itself and inserting into new chromosome locations -
Why is Alu classified as a retroposon?
Because it is though to require the retrovirus enzyme reverse transcriptase (rt) enzyme to make a mobile copy of itself. -
Give the scheme to explain how Alu elements transpose?
- First, the inserted Alu is transcribed into messenger RNA by the cellular RNA polymerase
- Then the mRNA is converted to a double-stranded DNA molecule by reverse transcriptase
- Finally, the DNA copy of Alu is intergrated into new chromosomal locus at the site of single - or double-stranded break
- First, the inserted Alu is transcribed into messenger RNA by the cellular RNA polymerase
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What is the function of th RNA polymerase III in each Alu element?
It is aninternal promoter toindependently initiate transcription of itself.
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DNA isolation and start-up PCR
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