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Agarose- Low EEO?

Agarose- Low EEO? - DNA Techniques

Agarose- Low EEO? - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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  #1  
Old 11-15-2006, 11:25 AM
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Smile Agarose- Low EEO?



Dear all,

Please help me out if can. What is the function of using a low electroendosmosis agarose gel in electrophoresis?

Thanks.
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Old 06-01-2008, 05:47 PM
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Default Re: Agarose- Low EEO?

good morning i am suresh, when i prepare 4% agarose (lowmelting) for RFLP i frequently faced the problem of bubbles. i tried hot plate, waterbath and microwaveoven to melt it. in all these bubbles were coming out. can any one help to solve this problem?
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Old 06-29-2011, 04:10 PM
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Default Re: Agarose- Low EEO?

"Electroendosmosis in electrophoresis refers to the flow of water under the influence of an electric field due to immobilized charge groups on the matrix, primarily sulfate and carboxyl groups. While the sulfate and carboxyl groups cannot migrate, their counter ions do migrate, moving toward the cathode, causing a net flow of water through osmosis in that direction. In the past, this problem often led to badly smeared bands."

Summary

Low EEO means less smeared bands
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Old 01-16-2014, 10:04 AM
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Default Re: Agarose- Low EEO?

Method for determination of EEO and protein mobility
For to evaluate the good performance of the electrophoresis system (buffer-agarose) employed for electrophoresis separation is need to know two fundamental parameters : electroendosmosis and protein mobility. We analyzed this parameters through a easy procedure, where human albumin serum (HSA) labeled with bromophenol blue and vitamin B12 are used. HAS is a negatively charged protein that fast migrate without hindrance in anode direction, while B12 molecule is a neutral substance transported only by electroendosmosis (EEO) that cathode migrate. For the test, a mixture of the two substances is placed on agarose gel connected with run buffer and a set up constant electric current is supply for a fixed time. After electrophoresis the distance that each migrated component from the starting point is measured, and these registered data are used to calculate the evaluation index.
EEO = cathodal migration distance of vitamin B12 from the well. Acceptable value between 5.0 and 5.5 mm;
Apparent Protein Migration (APM) = anodal migration distance of albumin from the well. Acceptable value between 7.0 and 7.5 mm;
Total Protein Migration (TPM) = EEO + APM. Acceptable value between 12 and 13 mm ;
Migration Index (MI) = EEO/TPM. Acceptable value between 0.40 and 0.45
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