No, definitely do not use normal melting point agar as you will denature the DNA. You use low melting point agar as it melts near a physiological temperature that will not denature proteins, nucleic acids and such. If you use normal melting point agar and raise the temp you will likely get false positives too.
Hope that helps!
First you layer normal agar on the slide and then mix the cells with low melting point agar when it temperature is close to 40 or less and layer it over the normal agar, it will give you better results.
No, there will be problem in maintaining DNA stability while using NMPA as Dr ai Benjamin Paul mentioned. To have better result go for prescribed chemicals which prepared under standard protocols.
NMPA (1.5%) and LMPA (0.75%) will give good result and try to avoid giving third LMPA layer.
NMPA layer on the slide acts a very good bed for adhesion of overlaid LMPA layer. Since, LMPA starts to melt even at a lower temperature, this semisolid layer acts as a suitable media for electrophoretic migration of DNA fragments through the gel layer.
Low melting agarose is melts at low temperature with fine quality powder and higher price incomarision with normal agarose Low melting agarose is very much suitable for comet assay. You can use also Normal agarose also.
all the best
low melting agarose is the best choice as it can gives god results and avoide false positive results which happen when use normal melting one as in this condition(normal melting) increase temperature may cause damage to the nucleic materials and so false positive.
best wishes
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