The plasmid was cut with an enzyme that cuts 2 times in the insert. This results in two DNA fragments with 1306 BP and 5497 BP.
Thus, the integration of the insert into the plasmid worked when two bands are visible.
Why are the DNA fragments of the different samples in the gel at different heights?
Sample 5-8 is higher than 1-4?
LV: uncut plasmid
1-12 cut plasmid
First of all it is clear that you have at least 3 different types of plasmid in this gel. Lanes 2-4 and 9-10 have one size insert and 5-8 have a different size insert. Secondly the plasmid backbone is different sized, compare 2 with 4 and 9 with 10. I don't think this is any sort of issue with the restriction digest but rather you just have different plasmid species. But without knowing the details of the experiment it is hard to hypothesize why.
I have checked the plate and I want to totally agree with Michael J. Benedik,
Hi there,
Apart from stating you have different plasmids and also some partial digest patterns (more than 2 bands) it's hard to conclude whether the expected construct is present on you gel without details of the DNA ladder for band size estimate.
The smallest fragment agrees with the marker. It is almost like saying the large fragment is almost uncut- which it truly seems the same as the large fragment is almost the entire plasmid. I am not sure if being such a large fragment it might be tangled, like a duplex and not a traight thread. The star activity and the conditions for the assay?...
How does the enzyme cut twice? (It cuts and recombines? I supose it does not mean blunt ends?)
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