need some help Gel Electrophoresis Calculation

[Skhan]

I am currently a student at a university and needed some help!

Ive got a practicle in which

restriction enzyme digest of plasmid DNA. Electrophoresis of PCR product, native and digested plasmid DNA


i need help in calculating ive got the results for the practicle if some one could guide me in brief what i have to do

i do remember parts that you have to Log10 the results


1kbp Marker
1= Plasmid DNA
2= xhol1 cut
3= BamH1 and Xhol 1 Cut
4= PCR Plasmid
100bp Marker
thank you !

[admin]

Hello,
I am not sure I understand your question. Are you trying to size the bands in marked 1-4? Do you know what 1 kb marker it is or the sizes of the marker bands in lane 1kb and/or 100bp?

cheers

[butters]

u need to rerun the gel so the distance for dna 1, 2 and 3. for ur pcr plasmid u need ~2% gel. Distance of the markers are too near.

haha, if u can supply the exact size for the marker bands. admin will show u

[Skhan]

sorry guys the results we did where not correct soo i have the results which the lecturer has posted !

attached are the results and also explaination if anyone could guide me to calculation

[Skhan]

heres the attachment

[Skhan]

can any one help out ? or just give me a hint where to start

[DeniseM]

To size bands on an agarose gel, measure the distance from the well to each band in the KB ladder (it doesn't matter if you measure to the top, middle, or bottom of the band as long as you do the same to each band). Plot the distances against the known value of the band on semilog paper, and draw a best fit line. Measure the distance from the well to the unknown sample, the same way you did for the ladder, and use the graph to determine the size. Find the distance, and draw a straight line up until it meets the best fit line. Draw a line straight over from where the intersection of the best fit line to determine the size. You will have trouble with lane 4 and the 100 bp ladder because the ladder doesn't have good enough resolution to distinguish between the bands.

Hope this helps,
Denise