Why needing normalization technique?

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Discussion Overview

The discussion revolves around the need for normalization techniques in experiments, particularly in the context of microarrays and transfection. Participants explore the reasons for normalization and seek additional resources for understanding the topic better.

Discussion Character

  • Exploratory
  • Technical explanation
  • Conceptual clarification

Main Points Raised

  • One participant questions the necessity of normalization techniques in experiments, specifically in microarray and transfection contexts.
  • Another participant explains that normalization is needed to account for systematic errors, such as differences in dye labeling efficiencies, scanning properties, and transfection efficiencies.
  • A request for general discussions or useful links about normalization techniques is made by a participant.
  • A participant provides a link to a resource on microarrays and lists commonly used normalization procedures, including Total Intensity normalization and LOWESS Normalization.
  • One participant expresses uncertainty about the terms "microarrays" and "transfection," asking for clarification.
  • Another participant clarifies that microarrays are used to measure gene expression through hybridization of RNA to probes, while transfection refers to introducing DNA into eukaryotic cells for gene expression measurement.

Areas of Agreement / Disagreement

The discussion includes varying levels of understanding about normalization techniques, with some participants providing explanations while others seek clarification. No consensus is reached on the necessity or methods of normalization.

Contextual Notes

Some participants express uncertainty about specific terms and concepts related to normalization, indicating a potential gap in foundational knowledge that may affect the discussion.

mountain
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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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