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Gene

The document provides an overview of gene chip technology, specifically DNA microarrays, which are used to measure gene expression across thousands of genes simultaneously. It discusses the principles, types, applications, advantages, and disadvantages of DNA microarrays, highlighting their significance in genomics and personalized medicine. Despite their high costs and complexity, DNA microarrays are pivotal in advancing genetic research and diagnostics.

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10 views16 pages

Gene

The document provides an overview of gene chip technology, specifically DNA microarrays, which are used to measure gene expression across thousands of genes simultaneously. It discusses the principles, types, applications, advantages, and disadvantages of DNA microarrays, highlighting their significance in genomics and personalized medicine. Despite their high costs and complexity, DNA microarrays are pivotal in advancing genetic research and diagnostics.

Uploaded by

Marwa
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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GENE CHIP TECHNOLOGY

PRESENTED BY:
Natasha Irum
Shama Rashid
Sawaira Hayat
Afsheen Ara
Kainat Khan
Nitasha Irum
M.Phil. 1st Semester (BIOCHEMISTRY)
Contents:
 Introduction (By Shama Rasheed)

 Principle (By Afsheen Ara)

Steps involved in DNA microarray

Requirements of DNA MICROARRAY Technique

 Types of DNA chips and Applications (By Sawera Hayat)

 Advantages and Disadvantages (Kainat Khan)

 Conclusion

 Reference
Introduction:
 DEFINITION: A DNA microarray is a laboratory tool used to
measure the expression of thousands of genes at the same
time.
 It consists of a small chip or slide with tiny spots, each containing
a DNA sequence that corresponds to a specific gene.
 By applying a sample of labeled DNA or RNA to the array,
researchers can see which genes are active or inactive in a
particular cell or tissue based on how much the sample binds to
each spot on the chip.
 There are several synonyms of Gene chip such as DNA
microarray, DNA chips, DNA arrays, gene arrays and biochips.

 HISTORY: the use of miniaturized microarrays for gene expression


profiling was first reported in 1995, and a complete eukaryotic
genome (saccharomyces cerevisiae) on a microarray was
published in 1997.
Principle:
 The principle of DNA microarray technology is
based on the hybridization of labeled DNA or RNA
samples to a solid surface containing thousands of
known DNA sequences.
 It is widely used in genomics and molecular biology
to monitor gene expression, detect mutations, and
identify genetic variations.
 MIAME ( Minimal Information About a Microarray
Experiment:
 It is a internationally adopted standard for the
designing of micro array experiment .
 Steps involved are :
continued..
 1.Experimental design : the set of hybridization experiments as whole

 2. Array Design: A DNA microarray consists of a grid of small spots, each


containing a specific DNA sequence, often referred to as probes. These
probes are short segments of DNA that are complementary to the
sequences of interest, such as genes or specific genetic markers.

 3. Sample Preparation: The first step involves isolating the mRNA from the
sample cells (for gene expression studies). The mRNA is then reverse-
transcribed into cDNA (complementary DNA) and labeled with fluorescent
dyes (e.g., Cy3 and Cy5).

 4. Hybridization: The labeled cDNA (which represents the gene expression


in the sample) is then applied to the microarray. The cDNA will hybridize
(bind) to the complementary DNA probes on the array. The hybridization
occurs due to base pairing between complementary nucleotides.
 5. Washing after hybridization: The microarray is
washed to remove unbound or non-specifically
bound DNA. This ensures that only specific
hybridizations are detected.

 6. Detection: The microarray is scanned using a laser


or fluorescence scanner to detect the fluorescence
emitted by the labeled cDNA. The intensity of the
fluorescence at each spot corresponds to the
amount of hybridization, indicating how much of the
specific gene or transcript was present in the sample.

 7. Data Analysis: The data from the microarray scan is


analyzed to identify which genes are actively being
expressed in the sample, their relative expression
levels, and any differential gene expression between
different conditions or samples.
 Software: Some common software used for DNA
microarray analysis include Affymetrix, Agelint and
illumina’s Genomestudio.
Requirements of DNA MICROARRAY
Technique:
 There are certain
requirements for designing a
DNA microarray system:
❑ DNA CHIP
❑ Target sample (Fluorescently
labelled)
❑ Fluorescent dyes
❑ Probes
❑ Scanner
Types of DNA microarray:
 Main types of micro arrays used commonly are :
 DNA Microarray :
 In this type we take DNA as sample to analyze.
 Binding mutations are identified through this type.
 Aiding disease research.
 DNA microarrays detect mutations in a gene's DNA,
helping diagnose diseases and genetic disorders.
 Dna microarray is further classified into two mains
those are :
 cDNA micro array
 Oligonucleotide array
 cDNA micro array :
 Most common type
 Follows the basic technique
 Involves production or mRNA then cDNA and rest of the steps discussed obove

 Oligonucleotide array:
 An oligonucleotide micro-array works in the same way as a cDNA microarray,
 but instead of using longer cDNA sequences, it uses shorter sequences called
oligonucleotides .
 These oligonucleotides are shorter pieces of DNA that match specific genes.
 Protein Microarray:
 Protein microarrays track protein
interactions
 They focus on protein functions and
their roles in cells.
 providing insights into gene
expression not fully captured by
DNA microarrays.
 Peptide Microarray:
 Peptide microarrays are similar to
protein microarrays but focus on
mapping antibody binding sites
and protein interactions.
 They help in studying diseases and
developing vaccines and
diagnostic tools.
Applications of DNA Microarray:
 In humans, they can be used to determine how particular diseases affect
the pattern of gene expression in various tissues, or the identity of the
infecting organism. Thus, in clinical medicine alone, DNA microarrays have
huge potential for diagnosis.
Besides, it has applications in many fields such as:
➢ Discovery of drugs
➢ Proteomics
➢ Functional genomics
➢ Diagnostics and genetic engineering
➢ DNA sequencing
➢ Gene expression profiling
Advantages of DNA Microarray :

 Provides data for thousands of genes in real time.


 single experiment generates many results easily
 Fast and easy to obtain results.
 Promising for discovering cures to diseases and cancer.
 Different parts of DNA can be used to study gene expression.
Disadvantages of DNA Microarray:
 Expensive to create: The cost of DNA microarray can range from few
thousand to hundred thousands of dollars, depending on the complexity of
the design, the numbers of probes.

 The production of too many results at a time requires long time for analysis,
which is quite complex in nature.

 The DNA chips do not have very long shelf life: physical and chemical
degradation, handling etc
Conclusion:
DNA microarrays have revolutionized our understanding of genomics and
gene expression. Despite their limitations, they provide a powerful and efficient
way to analyze large-scale genetic data.

Future Outlook: As technology continues to evolve, DNA microarrays will


become more accurate, affordable, and accessible, helping to advance
personalized medicine, cancer therapies, and genetic research.
References:
 https://www.slideshare.net/Haddies/dna-microarray-dna-chips
 https://www.slideshare.net/mahendrareddy59/dna-microarray-56074935
 https://www.news-medical.net/life-sciences/DNA-microarray.aspx
 https://www.researchgate.net/figure/Steps-in-DNA-Microarray-Experiment-
III-GENE-EXPRESSION-DATA-A-typical-DNA-microarray_fig1_236868392

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