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Genome biologyGenome biology
TopicsTopics1.1. DefinitionsDefinitions
2.2.The structure of the genomeThe structure of the genome
3.3.The function of the genomeThe function of the genome
4.4.Methods of genomicsMethods of genomics
1.Definitions1.Definitions
Definitions- 1Genome: definition 1. The information coded in the material of inheritence of an organism
definition 2. The haploid DNA (of a cell) of an organism
1. Nuclear genome 2. Mitochondrial and chloroplast genomes
Transcriptome: 1. Full transcriptom: - the total amount of mRNAs of an
organism 2. Cellular transcriptome:- the total amount mRNAs of a cell of an
organism in an experimental situation
Proteome: 1. Full proteome: - the total ammount of proteins of an organism 2. Cellular proteome: - the total ammount of proteins of a cell in an
experimental situation
Definitions- 2Genomics (genome biology)
1. Structural genomics, def: genetic mapping and comparison of individuals
a. determination of the genomic sequence (human, mouse, chimpanzee, etc.)b. genome variability: intraspecific polimorfismc. genome evolution: interspecific polimorfism
2. Functional genomics:def 1: examination of the transcriptomedef 2: examination of the function of the
genes
2/1 Functional genomics-I: transcriptomics 2/2 Functional gemomics-II: proteomics
Scope: - collecting of cDNAs - measuring the differentions in mRNA expression: transcriptomics - measuring the differentions in protein expression : proteomics
Definitions- 3
Alternative grouping:
• Genomics
• Functional genomics (transcriptomics)
• Proteomics
Definitions-4 Other „omics”:
• Phosphorylomics:The interaction between kinases and their substrates
• Metilomics: The methylation markings of the full DNA (3-5% in
mammals)
• Metabolomics: The interactions between enzimes and their substrates
• involved in metabolism
• Interactomics: Interaction between genes
• Lipidomics: The collection of lipids
Omics: system biological approach
The phosphorylome of the yeast
Red dots: kinasesBlue dots: substratesGreen lines: connections
Metabolome
2. The structure 2. The structure of the genomeof the genome
2a. The structure of the DNA
2b. The variation of the DNA
2c. The evolution of the DNA
2a.The structure of the 2a.The structure of the DNADNA
Genome programs
The human genome
Genome programs- history
1990 The genomes of the viruses
1995 The first prokaryotic genome – H. influenzae
1996 The first eukaryotic genome – yeast
1998 The first multicellular genome – C. elegans (string worm)
2000 Drosophila melanogaster, Arabidopsis thaliana (goose-weed)
2001 Human genome: draw version (90%): 30-35,000 gene
2002 Mouse genome: draw version
2004 Human genome: full version (99%): 20-25,000 gene
2005 Chimpanzee genome: draw version
Genome programes- active ( 300)
a. Non mammals: Lot of viruses: small genomeE. coli: model organismOther bacteria: H. influenzae, etcAmoeba: different genomeString worm (C. elegans): model organism Fruit fly: model organismBee: livestock, intelligent insect3 wasp speciesTripanosoma + malaria gnat: health care Triboleum castaneum: pest, modell animal of beetlesSea star: modell animal Goose-weed (arabidopsis) modell, rice + coffee: agriculture
b. Mammalshuman: vanity, self-study, health caremouse, rat: model organismbovine: livestockdog: huge number of genetic variants, homogenic races (in-bred breeds)chimpanzee: relativeorangutan: Rhesus monkeyWallaby (kanguru)Marmoset (monkey)
Genome programs – the competition
director, NIH National Human Genome Research Institute
Craig Venter
Francis Collins
Bill Clinton
Head of the Celera Genomics
President: USA
The set-up of the human genome
–What did they found?I. Not 100,000 - 150,000 genes, but: 20,000 - 25,000 - barely more, than fruit fly and the C. elegans, but the proteome is ~10x
as big
II. The bigger part of the genome is non-coding: waste - or selfish DNA? –maybe functional?
III. Nearly all insect and string-wormal genes are inside of us as well.
IV. This is not true conversely: immunity genes: antibodies, MHC, cytokinines; apoptotic genes, etc.
V. Numerous proteins from one gene: a human gene codes for an avarige of 2,6 protein: alternative splicing
VI. More transcription factors
VII. Huge enhancer region
VIII. More complex domain structure
The human genome
Total genome45% Transposable elements 25% introns + UTR20,7% other intergenic sequences5%
Simple repeats (microsatelites; VNTR-s)
Protein coding sequences
large duplications
3%
1,2%
21% LINE 13% SINE 8% 3%
Non-LTR retrotransposonsLTR - retrotransposons
DNA transposons
53% repetitive sequences
LTR:long terminal repeat (regulatory role)LINE: long interspersed elements;SINE: short interspersed elements:
Total genome45% Transposable elements 25% introns + UTR5%
21% LINE 13% SINE 8% 3%
Retrotransposons DNS transposons
LTR retrotransposones(Retroviruses, and other functioning retroposons) (450,000 copies)
Non-LTR retrotransposons(degenerated viral genes)(850,000 LINE, 1500,000 SINE)
Protein coding sequences
1,2%
„Copy and paste”
„cut and paste”
The human genomeSimple repeats
3%
LINE: long interspesed elements; SINE: short interspersed elements:
20,7% other inter-genic sequences
large duplications
The human genome -transposable elements
CP NC Pr RT RNáz H Int
gag pol env
LTR capsid nucleocapsid proteinase ribonuclease H envelope LTR
Reverse transcriptase integrase
CP NC Pr RT RNase HInt
gag pol
RT RNase H
gag? pol
A B
ORF
transposase
I. class
II. class
Retroviruses (1%)
SINE-s (Alu)
Retrogenes
DNA transposons
LTR retro-transposones
polyA
polyA
polyA
LINE-s (pl. L1)
I. Class: retotransposons I/1. LTR transposons I/2. Non-LTR transposons II/21. LINE-s II/22. SINE-s II/23. Retrogenes
II. Class: DNA transposons
3%
33%
IR IR
LTR: long terminal repeat
LTR LTR
7%
The human genome-retroviruses
CP NC Pr RT RNase HInt
gag pol env
LTR capsid nucleocapsid proteinase ribonuclease H envelope LTR
Reverse transcriptase integrase
LTR: long terminal repeat
gag: capsid (structural elements)pol: polimerase: reverse transcriptase, integrase, proteinase, RNase H env: envelope (structural elements)
Low copy number (10-1000 copies) human endogene retroviruses are presentin 1% of the genome
The human genome- Retroviral infection
The human genome - retrotransposons
CP NC Pr RT RNaseHInt
gag pol
RTRNase H
gag? pol
A B
ORF I. class
SINE-s (Alu)
Retrogenes
LTR retro-transposons
polyA
polyA
polyA
LINE-s (pl. L1)
LTR retrotransposones: from human endogene retroviruses, 10 – 1000 copies
LINE-s: in human L1 is the most common; present in 100,000 copies, but Lots of them are degenerated pseudogenes (non perfect reverse transcription).The 3,500 full length (6,1 kb) L1 –s 1% have promoter and two intact ORFs. LINE mobilisation in germ line and somatic cells as well.
SINE-s: 500,000 – 900,000 Alu copies (the most succesful transposone in human). All Alu element was created from a 280 bp + polIII promoter containing 7SL RNS gene. AluI restriction enzyme recognition sites are present in them.
Reverse Transcription!
LTR LTR
I. Class: retotransposons I/1. LTR transposons I/2. Non-LTR transposons II/21. LINE-s II/22. SINE-s II/23. Retrogenes
II. Class: DNA transposons
The human genome - DNA transposons
transposase II. class
DNA transposons
DNA transposons:
- The transposase responsible for the flip: how does it multiply?- More than 60 families: Charlie, mariner, Tigger, THE1, etc -The mariner family is similar to the the transposones present in insects: horizontal gene transfer?
IR IR
IR: inverted repeat
21% LINE 13% SINE 8% 3%
Retrotransposons DNA transposons
I. Class: retotransposons I/1. LTR transposons I/2. Non-LTR transposons II/21. LINE-s II/22. SINE-s II/23. Retrogenes
II. Class: DNA transposons
Total genome45% Transposable elements 25% introns+ UTR5%
Simple repeats (microsatellites; VNTR-s)
Large duplications: minisatellites and macrosatellites
3%
The human genome - microsatellites, minisatellites, macrosatellites
Microsatellites: small 4 base pair long or shorter repeats: 1 – 15 kilobasepairs long-CA/TG repeats in the 0.5% of the genom – yet their function is not known, „replication slippage” - AAAAs and TTTTTs- trinucleotid repeats CAA (Glu), ACA (ala): neuronal disorders; transcription factors in dogs
Minisatellites: 1 – 15 kbs repeats: like telomer: 15 kb: TTAGGG hexamer -the telomerse forges to the end of the chromosomes
Macrosatellites: several hundred kbs repeats
Exons
1,2%
Satellites: highly repetitve sequencesDuplications: importance in evolution
DNAsatellites
20,7% other inter-genic sequences
Total genome45% Transposable elements 25% introns + UTR5%
Simple repeats Protein coding sequences
Large duplications 3% 1.3%
The human genome - exons and introns
Exons: - protein coding DNA sequences + UTRs
Introns: - cut out- alternative splicing, other alternative processes.- are they functional?
leaderleader
E1E1
I1I1 E2E2 I2I2 trailertrailer
E3E3
Coding sequenceAUG Stop
pre-mRNS
polyA signal
5’-UTR 3’-UTR
20,7% other inter-genic sequences
Total genome45% Transposable elements 25% introns + UTR20,7% other inter-genic sequences5%
Simple repeats
Protein coding DNA sequences
Large duplications
3%
1.3%
The human genome- other intergenic sequences
1. Unidentifyable degenerated transposones2. Pseudogenes: 2 types (reverse transcripted RNA, duplicated DNA)3. Regulatory elements: promoters, enhancers, silencers4. others
2b. The variability of 2b. The variability of the DNAthe DNA
- intraspecific variability- intraspecific variability
Human genom diversity programes
The genetic code of the phenotypic variability – coding vs regulatory sequences
Human genome diversity programs
-From 1990 programs to map the polimorfism of the human genome. Importance: genealogic, medical
-From 2005 Genographic Project (National Geographic)
-mtDNA-Y chromosome
Genetic markers
The practicability of the data:
- Two theories on the origin of Homo sapiens (From Homo erectus):
multiregional theory – African origin (mitochondrial Éva)
- The wanderings of modern man.
Inheritence
somatic chromosomes
Inheritance
somatic chromosomesY chromosomeMitochondrial DNA
Inheritance
somatic chromosomesY chromosomeMitochondrial DNA
Genes STR*-s
Genetic markers on the Y chromosome
STR: short tandem repeats
16,569 nukleotide
Genes on the mitochondrial DNA
Hiper variable region
„Common” origin„Common” origin
100.000 years ago
Homo sapiens Homo sapiens Homo neanderthalensis
Homo erectus African Homo erectus
Asia
n H
omo
erec
tus
European Homo erectus
1.8 million years ago
Hypotheses: Multiregional Origin Out of Africa------------------
√
67,000 yr
African originAfrican origin
13,000 yr40-60,000 yr
20,000 yr
130,000 yr
40,000 yr
Comparison of mitochondrial DNAs: winning of „Out of Africa” hypothesis over „Multiregional Origin” hypothesis.
yr
The genetic base of the phenotypic variability: coding vs regulatory sequences
1. Genes and proteins functional variance
2. The theory of neutrality
3. Intragenic variability in the regulatory regions
4. Variance in the coding region of the regulatory genes
Genes and proteins - functional variance
Traditional concept
The different gene-products are responsible for the phenotypic variance in efficiency and function
The theory of neutrality
The gene variants (alleles) are functionally the same !
Motoo Kimura
- The majority of gene substitutions are not responsible for amino-acid changes (sinonim changes)
- The vast majority of aminoacid substitutions do not changes the function of the protein (chemically similar aminoacid substitutions: conservative change)
- A The function of the genes did not changed through evolution, Gene variability do not cause phenotypic variability. These are true for the most genes.- except for defective genes
Of Mice and ManSignificant polimorfism in the regulatory sequences- variability: expression level and tissue-specifity
Intraspecfic variability in the regulatory regions
Intraspecfic variability in the regulatory regions
P gene A
P gene A
P gene A
P gene A
enhancers promotersindividuals
1
2
3
4
The gene regulation theory: variability in gene expression
Variants of gene „A” with identical function but differing controlling regions
Coding variance: number of triplet repeats (number of glutamine and alanine repetitions)
1931
1976
Q19A14
Q19A13
. . . CAACAAGCACAAGCAGCA . . .
Q Q QA A AQ: glutamineA: alanine
bull terrier
Harold Garner and John W. Fondon Revival of gene function theory:
Variability in the sequence of transcription factors
runx-2 gene
Intraspecfic variability in the regulatory regions
2c. The evolution of the 2c. The evolution of the DNADNA
- interspecific variability- interspecific variability
Differences between genomes
The chimpanzee in us
Exaples Size (bp) Length (m)
HIV-1 9.8x103 10-6
fage 4.8x104 10-5
T4 fage 1.7x105 10-4
E. Coli 4.6x106 10-3
Drosophila 1.8x108 10-1
Mouse 3.5x109 1Dog 3.4x109 1Horse 3.3x109 1Human 3.4x109 1Corn 5.0x109 1Lilie 3.6x1010 10Amoeba 2.9x1011 100
The genome size differs between species and is not in relation with phenotypic complexity
The differences between genomes - Genome size
The differences between genomes - Number of the genes compared with the total number of the cells
Number of the genes: number of the cells:
Human: 20 – 25,000 1014
Fruit fly: 13,500 -C. elegans: 19,100 959
Differences between genomes Sctructural and functional differences
DNA similarity:
Human - chiken: 60%Human – mouse: 88%Human – chimpanzee: >98%
Same proteins
Ascidian –human: 80%Human – fruit fly: 40%Fruit fly – human: 61%C. elegans – human: 43%Yeast – human: 46%
Same domains:
Human – fruit fly, C. elegans > 90%,Exon shuffling in human , 2x as much gene
Rather „copy and paste”, than „cut and paste” mechanism
4 theories about the differences:
1. Evolution of regulatory proteinsa. FOXP2 gene: mutation: disorder in speech and articulation; 2 amino acid differences between human and chimp b. ASPM and MCPH1 genes mutation: microcephaly; their expression levels are higher in neural precursor cells
2. Evolution of regulatory sequences A general increase in the expression level of genes in brain; it is difficult to detect it at the level of DNA
3. Retainment of juvenile characters Lack of body fur, higher brain/body weigh ratio, the form of out skull is similar to that of chimp kid
4. Neotenia theoryCompare to the rest of the body, the development of head accelerated
Human genome - chimp genome
1. Chromosome number: 23 vs. 24
2. Genetic alterations:
3. More Alu and L1 sequences in human (short repetitive sequences)
- Point mutations (complete genome)… 1.23%- Point mutations (coding sequences).... 1% - Duplications: ……………………….. 2.7%- Insertions, deletions:………………… 3.0%- Several Chromosomal rearrangements
The chimp in usThe chimp in us
3. The function of the 3. The function of the genomegenome
3a. Gene expression
3b. Genome expression
3c. An astonishing RNA world
cytoplasmcytoplasmnucleusnucleus
ERER
GolgiGolgi
pol-II
pre-mRNA
mRNApolyA
cap
DNADNA
protein
ribosome
RNA polimerase-II
Gene expression
Alternative gene usage - instrument of complexity
Alternative……
- …promoter-usage- …splicing- …polyadenylation- …gene expression
P1P1 Ex1Ex1
Coding region
pre-mRNA
mRNA
protein
TT
Alternative promoter usage
Ex2Ex2 Ex3Ex3I2I2I1I1Promoter 1 terminator
ribosome
Ex: exonI: intron
P2P2Promoter 2
Epidermal cells
pol
Coding region
pre-mRNA
mRNA
protein
TTEx1Ex1 I2I2I1I1 Ex3Ex3Ex2Ex2terminator
Ex: exonI: intron
Alternative promoter usage
P1P1Promoter 2
P2P2
Neuronal cellsPromoter 1
pol
Alternative RNA splicing
E2E2PP E1E1 E4E4
E1E1 E2E2 E4E4
E3E3E1E1 E3E3 E4E4
E1E1P1P1 E3E3 E4E4E2E2P2P2
E1E1 E3E3 E4E4
E2E2 E3E3 E4E4
E2E2PP E1E1 E4E4
E1E1 E2E2 E4E4
E3E3E1E1 E3E3 E3E3
PA2PA2PA1PA1
DNADNA
DNADNA
DNADNA
Alternative splicingAlternative splicing
Alternative splicing + promoter usage Alternative splicing + promoter usage
Alternative splicing + polyadenylationAlternative splicing + polyadenylation
gene 2
Nerve cells
Neuron-specific enhancer
P2 TEx2 Ex3 Ex4I3I2
Cell-specific gene expression
gene 1
P1 TEx1 Ex2 Ex3I2I1
Ex1 I1
Skin cell-specific enhancer
TF
gene 2
P2 TEx2 Ex3 Ex4I3I2
gene 1
P1 TEx1 Ex2 Ex3I2I1
Ex1 I1
Skin cell-specific enhancer
S
Neuron-specific enhancer
TF
Skin cells
Cell-specific gene expression: histone pattern!
1. Type of transcription factor expressed in a cell
2. The accesability of the regulatory region of a gene
The expression of the genome
Expression changes for a lot of genes:
1. Different tissues2. Between humans3. Diseased - healthy
Transcriptome
Astounding RNA world
1. The larger half of the genome is transcribed
- The ncRNA coding part of the genome is 50 times longer, than the part which encodes coding RNA (genome tiling arrays, cDNA cloning)
2. There are regulatory antisense RNAs everywhere
- a significant portion of the genes are under the control of trans-asRNAs (miRNAs):
1 miRNA – more gene; 1 gene – more miRNA (transcriptome analysis)
- a significant portion of the genes are under the control of trans-asRNAs (miRNAs): (EST analysis)
- Two discoveries
1. New functions of RNAs
- Traditional function: information transmission from DNA to proteins (mRNA), and conribution in this process (tRNA, rRNA)
- New functions:●RNAs are independent information carriers●RNAs regulate the expression of genetic information
2. Genetic regulation
- Traditional theory: the genetic regulation is achieved by theinteraction between transcription factors and the cis regulatoryelements (promoter, enhancer, silencer)
– Present theory: essential rule of regulatory ncRNAs
An astonishing RNA world - Two surprises
coding sequences non-coding sequences non-coding/coding Species protein-coding gene genome size MB % MB % sequence ratio (db) (MB)Complete genome Human 20-25 000 2851 34 1,2 1619 57 47,5 : 1 Mouse 20-25 000 2490 31 1,3 1339 54 42,5 : 1 Fruit fly 13 500 120 22 18 53 44 2,4 : 1 C. elegans 19 000 100 26 26 33 33 1,3 : 1
Non-repetitive sequences Human 1455 33 2,3 867 60 26,1 : 1 Mouse 1422 29 2,0 811 57 28,5 : 1 Fruit fly 109 21 20 48 44 2,2 : 1 C. elegans 86 25 29 26 33 1,1 : 1
Coding – non coding
Types of RNAs
Types of RNAs
RNAs
Coding RNA Non-coding RNA
mRNA tRNA rRNA aoRNA miRNA snRNA snoRNA
Transcription RNA
siRNA
Regulatory RNA
messenger transfer ribosomal small interfering micro small nuclear
antisense overlapping small nucleolar
33.
a. trans-antisense RNAs: imperfect homology
b. cis-antisense RNAs: perfect homology
gene
antisense RNA
cis trans
DNA
Regulating antisense RNAs
Micro RNAs
Micro RNAs
nucleus
Droshatranscription
pri-miRNA
pre-miRNA
DICER
exportin-5
RISC
mature miRNA
blocked mRNA
1 23
4
5
cytoplasm
(trans-antisense RNAs)
Discovered in 2000
34.
degradation translation block
pre-miRNA
miRNA
The mechanisms of miRNA action
The mechanisms of miRNA action
The function of miRNAs: - Ontogenesis (timing), apoptosis, cell proliferation, oncogenesis
35.
cis-antisense RNAs - overlapping antisense RNAs
5’3’DNA
mRNA
5’ 3’
5’ 3’
coding strand
non-coding (antisense) strand
over-lapping
RNAs
5’3’5’ 3’
5’3’5’ 3’
5’3’5’ 3’
Complete overlap
Convergent
Divergent
5’3’5’ 3’
5’3’5’ 3’
1. Blocking of tranlation elongation
3. RNA interferencie (?)
RISC
RNáz RNáz
ribosome
FUNCTION
5’3’5’ 3’
FactorFactor: translation, regulation of half-life
2. Blocking of translation initiation
Overlapping RNAsOverlapping RNAs ((ciscis-antisense -antisense
RNAs)RNAs)
4.Tools of Genomics4.Tools of Genomics4a. The structure of the DNA 1. cloning – genome library
construction 2. sequencing
4b. The expression of the DNA
1. cDNA library, EST library 2. DNA mikroarrays 3. Protein arrays
4c. Bioinformatics
Gene libraries
Genom library: collection of clones, in wich every pieces of the genome of a particular organism can be found.
Usage: sequencing (genome projects), isolation of genes.
cDNA library: (cDNA: copy DNA) The cDNA library contains a cDNA copy of each mRNA of an organism (tissue or cell type). It represents the transcriptome.
Usage: gene structure determination, isolation of cDNSs (intronless gene).
EST library: ‘Expressed Sequence Tag’: either the 5’ or the 3’ end of a cDNA. It also represents the transcriptome of an organism (tissue or cell type), however, because of the smaller clone sizes, it can be handled, sequenced easier and faster.
Usage: determination of the transcription of a cell or tissue type (What kind of genes are expressed in this tissue?).
EST libraries were used for the rapid sequencing of the „active” genome.
VectorsSeveral different vectors can be used for theconstruction of gene libraries, like:
PlasmidBacteriophage (lambda)CosmidBAC (Bacterial Artificial Chromosome)YAC (Yeast Artificial Chromosome)
Vector Maximal clone size Numb. of clones requiredfor a complete library
Plasmid 10 Kb 107
Bacteriophage 20 Kb 5 x 105
Cosmid 45 Kb 2 x 105
BAC 500 Kb 5 x 104
YAC 1 Mb 104
Structure of vectors
Plasmid: replication origin, antibiotic resistance gene, multi cloning site with many unique restriction sites.
BAC: Bacterial Arteficial Chromosome. In fact, it is a plasmid, with a replication origin, that can be found on native giant plasmids (like F-factor).
Construction of genome librariesFragmenting the DNAThe aim is to produce overlapping DNA fragments, that have an appropriate lenght, according to the vector content. We can use restriction endonuclease with 4pbs long recognition site, or we can physically shear the DNA (e.g. pressing through a thin capillary).The animation shows, how the overlapping DNA fragments are generated by partial digestion with an endonuclease.
1 2 3 4 5 6
Constructing genome libraries2.: ligation into plasmid vector1.: partial digestion with restriction endonuclease
1
2 53 4 6
1
2 5
3
4
6
3.: transforming into E. coli
3
1
2
4
5
6
3
1
2
4
5
6
Constructing genome libraries2.: ligation into plasmid vector
Screening of gene librariesHybridization
5’ 3’
5’3’G T G C A C
5’3’G T G C A C
C A C G T G
probetarget
Southern blotAgaroze gel electrophoresis
transfer
Nitrocellulose or plasticmembrane
Hybridization with a labeledDNA probe
Screening of gene librariesColony hybridization
lysis
transfer onto membrane
hybridizationwith a labeledprobe
cDNA-librariesHow can we produce cDNA?
AAAAAAAAA 3’mRNA 5’
TTTTTTTTT 5’3’cDNSfirst strand
3’ CCCCC
5’ GGGGGcDNASecondstrand
3’
1. RNA (mRNA) purification: we can use total RNA or mRNA extract
2. Reverse transcription: by using of oligo dT primers and reverse transcriptase (RNA-dependent DNA polimerase) the first strand of cDNA is synthesized
3. RNase treatment
4. Linker synthesis: the terminal deoxinucleotidil transferase (DNA polimerase, which doesn’t require any template) adds the C linker to the 3’ end
5. Second strand synthesis: oligo dG primers are added and the DNA polimerasesynthesizes the second strand of cDNA.
The DNA chip (microarray)
It is for measuring the expression pattern of a large number of genes at the same time. In fact, the DNA chip is an inverted Southern blot: the known probes are covalently bound onto the glass surface. One chip contains 6-10000 gene specific probes. There are:
-cDNA-oligonucleotide and-sequencing chips.
1. Preparing the chip:- printing- in situ synthesis
2. Collection of tissue samples
control treated
3. RNA purification
4. Reverse transcription (fluorescent labeling)
5. Hybridization 6. Reading
The DNA chipReading and evaluation of data
Protein chipHow does it work?
control treated
Protein purification
labeling
Immune reaction
detection
Protein chipLabeling strategies
1. Direct labeling
2. Sandwitch method
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