Summary: Mboc Samenvatting
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1 DNA replication, sequencing and PCR
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Why is DNA replication considered semi-conservative?
Because every new double stranded molecule consist of one old and one new strand- the old strand is the template
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What is the chemical reaction behind DNA synthesis?
Theformation ofphosphodiester bonds whilehydrolysing thematching dNTP molecule of the template strand -
What are the frequency of mistakes?
- Without proof-reading: 1 in 10^5
- with proof-reading: 1 in 10^7
- with strand-directed mismatch repair: 1 in 10^10
- Without proof-reading: 1 in 10^5
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What is strand-directed mismatch repair?
When there is a error in the newstrand ,mismatch proofreading proteins will bind toDNA and detects error instrand . The error will be removed and theDNA strand will berepaired . -
What causes permanent changes in the template strand?
Chemical changes inDNA bases can causemutation in theDNA depurination : loss of purine bases (A andG ), will causedeletion in mutated stranddeamination :C -> U
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What are the main differences between DNA sequencing and PCR?
DNA sequencing determine the order of the bases, while PCR amply the DNA -
What is shotgun sequencing?
The method involves randomlybreaking up the genome into smallDNA fragments that aresequenced individually . Acomputer program looks foroverlaps in theDNA sequences , using them toreassemble thefragments in their correct order toreconstitute the genome.- works well for small genomes that lack repetitive DNA
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What are the difference between different sequencing platform?
Read length and error rate -
What are the advantages and restrictions of PCR?
Advantages: sensitive and fast
Restrictions: fragment length <20kb and homologous DNA sequences -
What are the techniques for detecting DNA polymorphism?
PCR-based: VNTR
PCR and sequencing-based/melting curve: SNP
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Translation of mRNA into proteins
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Gene cloning and manipulation
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Long and small non-coding RNAs
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Vesicular transport
