Hi, I am trying to find the number of basepairs in a plasmid from a gel electrop

Question

Hi,

I am trying to find the number of basepairs in a plasmid from a gel electrophoresis. The plasmid traveled 61 mm. When graphing the log of the base pairs in the marker lane versus the distance traveled. I found a calibration curve with an equation of y=-0.1369x + 4.4508. I plugged in 61 mm into x and solved for y which gave me y=0.532972494. I know in order to find the base pairs in the plasmid I have to take the inverse log of the value for y. However, I keep getting the wrong answer. Can someone find the correct answer of baspairs for the plasmid?

Thank you!

Explanation / Answer

I don't really get the question, but if you have an insert in a plasmid, and the you use primers in PCR, I would expect a segment with and without the insert to be "primed". Thus the insert is the larger minus the smaller band's base par number. I don't know in the case with EcoRI, though, it just "cuts". I assume that in this case the 500 bp segments represents the plasmid without the insert, while the 2000 bp represents the plasmid with the insert. On the other hand Gary has a point. If EcoRI is the same enzyme as was used during the insert, then the 500 bp segment must be the insert.

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