Is Low melting point agarose and agarose having low EEO the same? Is it possible to utilize agarose low EEO instead of low melting point agarose for COMET assay?
Hello Subham Chakraborty
No, you cannot use agarose having low EEO for COMET assay because the properties of agarose low EEO and low melting point agarose are different. Low melting point agarose has a low melting point due to chemical modification (≤65.5°C) as well as a low gelling point (24–28°C). The agarose will remain in a liquid state at 37°C. This will help to maintain an optimum temperature during the assay enabling recovery of intact DNA fragments after gel electrophoresis without denaturation or damage. The agarose with low EEO has a melting temperature of 88 ± 1.5 °C and a gelling temperature of 36 ± 1.5 °C.
You may pre-coat the slides with normal melting point agarose as it will increase the adhesion of the low melting agarose layer containing the cells for comet analysis. After having pre-coated the slides with normal melting point agarose, air-dry the slides overnight. When the slides have completely dried, you may add the mixture of cell-low melting agarose onto the normal melting point agarose-coated microscope slide.
Best.
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