Genetics 4 Flashcards
Analysis of nucleic acids - goal is what
To find defective genes or abnormal gene expression
Analysis of genes includes what
Locating a disease gene - require intense research
Diagnosing the presence of a disease gene - routine task
Analysis of gene expression is what
Quantifying RNA
Polymerase chain reaction (PCR) is used for
Amplification of DNA fragments
PCR can be used how
With allele specific oligonucleotides that bind only to mutant or wildtype alleles OR
Can be used to know if there are insertions or deletions by counting the length of the amplified fragment
Reverse Transcriptase PCR
PCR only works with DNA so if you want to analyze RNA you have to transcribe the RNA into DNA
Reverse Transcriptase PCR - In a first step…
Reverse transcriptase is used to transcribe RNA into cDNA
Reverse Transcriptase PCR - second step
cDNA serves as a template for a PCR reaction
Reverse Transcriptase PCR - the more RNA there is,
the more DNA will be amplified
Quantitative PCR - is what
Accurate quantification of specific DNA molecules
Amplification is monitored in real time
Quantitative PCR - Often used for
mRNA detection
copy of mRNA into cDNA first, then start PCR
Allele specific oligonucleotides are designed for what
To bind just one specific allele
Allele specific oligonucleotides - hybridization of genomic DNA to ASO demonstrates
presence of the allele
Allele specific oligonucleotides - what are the three possible outcomes
1 All of it binds to wildtype spot and non to mutant (homozygous for wildtype)
2 All of it binds to mutant and none to wildtype (homozygous for mutant)
3 Half to wildtype and half to mutant (your patient is heterozygous)
Sequencing
Reads it base by base and forms the nucleotide sequencing
Can be used to determine if a patient is heterozygous at a specific locust
