This document discusses genomic research technologies, including the invader assay. The invader assay is used to detect single nucleotide polymorphisms without requiring PCR. It works by using reporter and invader probes that are cleaved by a flap endonuclease only when the target sequence is present. The assay has the advantage of not needing PCR but has drawbacks of requiring a large sample size and having background reactions. Overall, the invader assay is a method for detecting SNPs and mutations without PCR amplification.
2. 2
We have got the map
(genomic sequence) to
find treasure so that we
can get treasure chest.
Key to open the chest
(Post genomic technology)
Getting treasure (new drugs) !!
Even if we get the treasure
chest (target gene), we can’t
open it (because we can’t
access to its function in
disease.)
Current genomic researches
have tried pulling out of all
nails on the chest.
However, the number of the
nails may be infiinite…
3. 3
What is the difference between human & ape?Genomic research
4. 4
Progress of Research
genome Genomic sequence・Polymorphism(SNP etc)
Transcriptome Gene transcription profile
Proteome Expression profile of proteins
Functional proteome
Metabolome
Genetic function
Post translational modification
Protein interation etc.
Time consuming and enormous cost
Elucidation of functional network of cellular molecules
5. 5
Technologies in each categories
Genome
Structure analysis(sequence)
Polymorphism analysis
-DNA chip -Invader assay
-Sniper assay -PROBE assay
-Luminex -PCR-SSCP
PCR-RFLP etc-
Transcriptome
・Differential expression analysis
・cDNA chip etc
Proteome
Identification
Protein function
Protein interaction
Ligand interaction
Post translational modification
・protein chip, peptide chip
・Y2H
・SELEX
・Phage display
・STABLE assay etc
6. 6
Used for
The detection of single nucleotide polymorphisms (SNPs)
Detection of mutations that confer resistance to antiviral
agents
Genotyping of isolates for epidemiological purposes;
categorizes patient isolates into 8 different HBV genotypes (A
to H) and 6 different HCV genotypes (1 to 6 with 24 subtypes)
7. 7
T
N
C
G
A
Reporter probes Flap FlapInveder probes
N
C T
G
N N
A
Cleavage Cleavage
Endoflap Nuclease
Fluorophore 1 Fluorophore 2
Quencher
Quencher
C
Cleavage
T
Invader Assay
8. 8
Advantage: PCR is unnecessary
Drawback: Quite large amount of sample is required
Background reaction exists