Microarray
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This document provides an overview of DNA microarray technology. It discusses the historical background beginning in the 1970s with Southern blotting and the development of microarrays in the 1980s. The key principles are that DNA microarrays allow analysis of thousands of genes simultaneously and efficiently through orderly arrangement of DNA sequences on a solid surface like glass. The main steps involve preparing the microarray slide through various methods, performing experiments with sample mRNA, fluorescence scanning, and data analysis to understand gene expression patterns. DNA microarray technology has wide applications in studying diseases, toxicology, and stem cell research.
Microarray
- 1. DNA Microarray Technology Under the guidance of Dr .Manjunath Ankitha Hirematha 3rd sem, MSc., Dept. of Biotechnology Kuvempu university.
- 2. CONTENTS • Introduction • Historical Background • Principle • Overview steps • Preparation of slide • Microarray scanning • Data analysis and normalization
- 3. DNA Microarray Technology To analyze the expression of thousands of genes in single reaction, very quickly and in an efficient manner. To understand the genetic causes for the abnormal functioning of the human body. To understand which genes are active and which genes are inactive in different cell types.
- 5. What is DNA Microarray Technology ? It is “an orderly arrangement of thousands of identified sequenced genes printed on an impermeable solid support , usually glass, silicon chips or nylon membrane”. DNA microarray chips are also known as DNA chips, DNA arrays, or biochips.
- 6. Historical background : Sir Edwin Southern • Southern blotting was developed in the year 1975. • The concept of DNA microarrays began in the mid 1980s. Sir Steve Fodor • Pin based robotic system was developed by Lehrach’s group in 1990. • Steve Fodor developed scanner for reading the output. •“Quantitative Monitoring of Gene Expression Patterns with a complementary DNA microarray” reported by Patrick Brown, Mark Schena and colleagues in Science (1995). •Mark Schena was proclaimed as the “Father of Microarray Technology”. Sir Patrick Brown Mark Schena
- 7. Principle of DNA Microarray:
- 8. An overview of steps involved in DNA Microarray
- 9. Preparation of DNA Microarray Slide : The solid supports used are glass, silicon or nylon membranes Length of slide is 25 X75mm Within the area of 3.6cm2 , 10,000 to 20,000 spots(genes) Diameter of spot is 50-150μm Distance between the spots is 200-250 μm. Fabrication • Photolithography • Robot spotting • Inkjet
- 10. Photolithography
- 11. Robot Spotting
- 13. Performing DNA Microarray Experiment Eg. Saccharomyces cerevisiae O2 O2 Centrifuge Supernatant is discarded Add extraction buffer
- 14. Remove that buffer containing mRNA and place in fresh tube AAAAAAAAAAA AAAAAAAAAAA AAAAAAAAAAA AAAAAAAAAAA AAAAAAAAAAA AAAAAAAAAAA Reverse Transcription TTTTTTTTTTTTT TTTTTTTTTTTTT TTTTTTTTTTTTT c DNA TTTTTTTTTTTTT TTTTTTTTTTTTT TTTTTTTTTTTTT Mix
- 15. ATGC ATGC ATGC GATC GATC GATC When When laser comes, laser comes, Merged image : TCAG TCAG TCAG
- 16. DNA Microarray Scanner • Fluorescent intensity is measured “Normalization means to adjust the microarray data for effects which arise from variation in the technology”.
- 17. Applications • Gene expresion profiling, SNP detection, human diseases etc.. • IPSC (Induced Pleuripotent Stem Cell) cell lines are validated and monitored. • Toxic studies
- 18. CONCLUSION Since this DNA microarray technology is used for the analysis of expression of thousands of genes at once, and has wide applications in in analyzing various diseases. It is one of the smartest technique.
- 19. REFERENCES •Tim Lenoir ,Eric Giannella (2006) The emergence and diffusion of DNA microarray technology. Journal of Biomedical Discovery and Collaboration(JBDC). 1:11 •Samuel K. Moore (2001) Biochips are now a critical tool for analyzing the human genome--and a lucrative product attracting technology giants. IEEE spectrum • http://www.premierbiosoft.com/tech_notes/microarray.html •http://www.genome.gov/10000533 •http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1590052/ •http://www.medscape.com/viewarticle/543871_2 •http://en.wikipedia.org/wiki/DNA_microarray#History •http://grf.lshtm.ac.uk/microarrayoverview.htm#m1 •http://www.ncbi.nlm.nih.gov/About/primer/microarrays.html •http://www.digizyme.com/portfolio/microarraysfab/photolith.html
- 20. Thank You