2024. 10. 26. 15:14ㆍLab skills
DNA microarray
: Microarray is a DNA chip in which very small DNA fragments are integrated on a solid surface
: Simultaneously measure the expression level of a large number of genes and compare the differences in gene expression between groups
*Analyze the expression patterns of multiple mRNAs transcribed from cells
: Since many samples are tested simultaneously, a large amount of data can be obtained in a short period of time
: Since it has various systematic errors such as dye bias/poor scanning/hybridization problems, correction is required in the analysis
Microarray Applications
a) New Drug Discovery
b) Diagnostics
c) Proteomics
d) Functional Genomics
m) DNA Sequencing
f) Gene Expression Identification
s) Toxicogenomics
Microarray experiment process
1) Sample collection
- Collection of Normal cells and Cancer cells to check gene expression
2) mRNA isolation
- Extract RNA from the collected cell lysate using a solvent such as phenol chloroform
- Remove rRNA and tRNA from the extracted RNA, leaving only mRNA
3) Labeled cDNA generation
- Perform reverse transcription of mRNA to generate complementary cDNA
- Use different fluorescent labels to distinguish between normal and diseased cells
4) Hybridization
- Synthesized cDNAs are spread on the chip and hybridization begins
- Unbound sequences are removed through washing
5) Instrument analysis
- Record cDNA fluorescence images using a microarray scanner
- The level of gene expression is measured based on the difference in fluorescence intensity for each spot
*Green: Normal
*Red: Gastric cancer
*Yellow: Common
Reference
1) https://www.khanacademy.org/science/high-school-biology/hs-molecular-genetics/hs-biotechnology/a/dna-sequencing
2) https://www.illumina.com/science/technology/next-generation-sequencing/ngs-vs-sanger-sequencing.html
3) https://ko.wikipedia.org/wiki/%EC%83%9D%EC%96%B4_%EC%97%BC%EA%B8%B0%EC%84%9C%EC%97%B4_%EB%B6%84%EC%84%9D
4) https://www.youtube.com/watch?v=0ATUjAxNf6U
5) https://www.semanticscholar.org/paper/Microarray-Analysis-and-Gene-Expression-%3A-A-Review-Subair-AlEisa/a75f33de77e9b7e220c102245fda6fb7bddf3245
6) https://warwick.ac.uk/fac/sci/lifesci/intranet/staffpg/support/genomics_new/microarrays/
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