Why needing normalization technique?

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Normalization techniques are essential in experiments like microarrays and transfections to mitigate systematic errors that can affect signal measurements. These errors can stem from various factors, including dye labeling efficiencies, scanning properties, and transfection efficiencies. Common normalization methods include Total Intensity normalization, LOWESS Normalization, Mean centering, Ratio Statistics, and Standard deviation regularization. Microarrays allow for the measurement of gene expression by hybridizing RNA converted to cDNA to probes on a chip, enabling analysis of numerous genes simultaneously. Transfection involves introducing DNA into eukaryotic cells, often used to assess gene expression by linking a gene promoter to a reporter gene. Understanding these concepts is crucial for accurate data interpretation in genomic studies.
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Why needing normalization technique??

Why do we have to normalize an experiment for instance in microarray or transfection?


Thanks.
 
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To account for differences in signal caused by systematic errors, like dye labelling efficiencies, dye scanning properties, power of the two lasers, print tip effects, spatial effects (for the microarray) and transfection efficiencies, cell survival, etc (for transfection).
 
I have searched for a general discussion about normalization, but could not find any good ones. Do you have any useful links?
 
Here is a link for microarrays http://www.dnamicroarrays.info/Data_Norm.html

The commonly used normalization procedures are Total Intensity normalization, LOWESS Normalization, Mean centering, Ratio Statistics, Standard deviation regularization.
 
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Stupid Question ( I seem to be good at asking them): what are microarrays and transfection?
 
Those are not stupid questions ;)

With a microarray you can measure gene expression. It is based on hybridizing the expressed gene products (RNA that has been converted into cDNA) to probes on a chip. When hybridization occurs you get a signal. In this way you can analyze all the genes in a genome (~30,000 in the case of humans).

Transfection is the method of introducing DNA into eukaryotic cells. It can also be used to measure gene expression, where a gene promoter is cloned in front of a reporter gene.
 

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