Why even 17th edition Harrison is important as far as entrance exam is concerned??? Genetics from Positive OHC

METHOD

PRINCIPLE

MUTUAL  DETECTED

Polymerase chain reaction (PCR)

Amplification of DNA segment

Expansion of triplet repeats, variable number of tandem repeats (VNTR), gene rearrangements, translocations; prepare DNA for other mutation methods

Reverse transcriptase PCR (RT-PCR)

Reverse transcription, amplification of DNA segment absence or reduction of mRNA transcription

Analyze expressed mRNA (cDNA) sequence; detect loss of expression

Traditional DNA sequencing

Direct sequencing of PCR products

Sequencing of DNA segments cloned into plasmid vectors

Point mutations, small deletions and insertions

METHOD

PRINCIPLE

MUTATION  DETECTED

Next-generation DNA sequencing

Sequencing of large contiguous genomic regions, exome of single or all chromosomes.

Deep sequencing for detection of mutations and SNPs. Sequencing of whole genomes of microorganisms

Microarrays

Hybridization of PCR products to wild-type or mutated oligonucleotides

Point mutations, small deletions and insertions

Genotyping of SNPs

METHOD

PRINCIPLE

MUTATION  DETECTED

Cytogenetic analysis

Unique visual appearance of various chromosomes

Numerical or structural abnormalities in chromosomes

Fluorescent in situ hybridization (FISH)

Hybridization to chromosomes with fluorescently labeled probes

Numerical or structural abnormalities in chromosomes

Southern blot

Hybridization with genomic probe or cDNA probe after digestion of high molecular DNA

Large deletion, insertion, rearrangement, expansions of triplet repeat, amplification

Polymerase chain reaction (PCR)

Amplification of DNA segment

Expansion of triplet repeats, variable number of tandem repeats (VNTR), gene rearrangements, translocations; prepare DNA for other mutation methods

METHOD

PRINCIPLE

MUTATION DETECTED

Reverse transcriptase PCR (RT-PCR)

Reverse transcription, amplification of DNA segment absence or reduction of mRNA transcription

Analyze expressed mRNA (cDNA) sequence; detect loss of expression

DNA sequencing

Direct sequencing of PCR products

Point mutations, small deletions and insertions

Sequencing of DNA segments cloned into plasmid vectors

Restriction fragment polymorphism (RFLP)

Detection of altered restriction pattern of genomic DNA (Southern blot) or PCR products

Point mutations, small deletions and insertions

Single-strand conformational polymorphism (SSCP)

PCR of DNA segment: Mutations result in conformational change and altered mobility

Point mutations, small deletions and insertions

METHOD

PRINCIPLE

MUTATION DETECTED

Denaturing gradient gel electrophoresis (DGGE)

PCR of DNA segment: Mutations result in conformational change and altered mobility

Point mutations, small deletions and insertions

RNAse cleavage

Cleavage of mismatch between mutated and wild-type sequence

Point mutations, small deletions and insertions

Oligonucleotide specific hybridization (OSH)

Hybridization of PCR products to wild-type or mutated oligonucleotides immobilized on chips or slides

Point mutations, small deletions and insertions

Microarrays

Hybridization of PCR products to wild-type or mutated oligonucleotides

Point mutations, small deletions and insertions

Genotyping of SNPs

METHOD

PRINCIPLE

MUTATION DETECTED

Protein truncation test (PTT)

Transcription/translation of cDNA isolated from tissue sample

Mutations leading to premature truncations

Pyrosequencing

Clonal amplification of single DNA fragments on microparticles followed by massive parallel sequencing

Sequencing of whole genomes of microorganisms, resequencing of amplicons

Multiplex ligation-dependent probe amplification (MLPA)

Quantification of PCR-generated amplicons reflecting the number of copies of a specific DNA sequence

Copy number variations