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DNA gel electrophoresis: protein contamination?

DNA gel electrophoresis: protein contamination? - Agarose Gel Electrophoresis Forum

DNA gel electrophoresis: protein contamination? - Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


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Old 04-24-2010, 02:12 AM
Pipette Filler
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Unhappy DNA gel electrophoresis: protein contamination?



Hi im kinda new here so if this is in the wrong section, then thats ok.


Well, what i an trying to find using 1.2% Agarose gel electrophoresis is to prove that there is protein contamination in my plant extracts (my plant extracts collect a month ago were freezed dried after collection before being grounded using a motar & pestle and finally was stored around 25 degrees (room temperature)).

This is so when i know there is protein contamination (which is what i want before finding some biophenols using High performance liquid chromatography.)

So what i did was to use 1ul microliter Dnase, rnase and proteinase K for each of my plant extracts. Positive control was the plant extract and was diluted (in 1/20) using distilled water. Negative control was Nuclease free water. Both contained 6x loading dye. A gene ladder was used in my sample to only know if the gel was working.

Note: None of the sample underwent purification process before loading the gel with the samples. Also note that the same samples tested on a biophotometer had a 230/260nm ratio of 1.01 for DNA/RNA purity and the 230 absorbance was low (about .8A) for the 1/20 sample


To get straight to the point, this is what each well contained

Well 1 : Empty
Well 2 : 3ul gene ladder with 2ul dye and 2ul Nuclease free water (NFW).
Well 3 : 4ul NFW and 2ul loading dye
Well 4: 7ul Diluted 1/20 sample and 2ul loading dye and 6ul of NFW
Wells 5-7 : 7 ul of diluted 1/20 sample with either 1ul of DNase, rnase and Pro K in each well, each well had 2ul dye and 6ul of NFW

well 8 had 1ul of DNase,Rnase and pro K (total 3ul) in the well with 6ul of diluted 1/20 sample, 2ul loading dye and 4ul of NFW

When the gel is made, only one band appear (which was the gene ladder), on the wells, and they were bright. However there is a faint brightness around the well. Could that mean something or that i have to make another agarose gel with the samples again?

Also if anyone can link my to a better protocol, that would be gladly accepted.
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contamination , dna , electrophoresis , gel , protein


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