Vectors and inserts were ligated after cutting with EcoR1 and Xho1, respectively.
The colonies were picked and cut again with restriction enzymes.
But after cloning, I saw the bands of the vectors appearing in different sizes.
I also checked before and after cutting vectors and inserts with the control band.
Why does the size of the vector change after cloning?
Hi Jihyun Park ,
I think your results are good, except the 6th one. My suggestion is, select any one good plasmid from 1 to 5 with high copy number (high density band) and confirm by sequencing it gene specific or universal primers. Best wishes.
the size of the vector has not changed after cloning for the first five of your clones which is evident when you compare it with your uncut vector. I guess you can take 1 of the clones and proceed for restriction checking with sites in vector and insert to confirm the backbone. also, you can run a ladder to check the size of the vector. Reconfirm with sequencing
In the case of 1-7 samples, it was cut and loaded with the restriction enzyme of the vector.
So I think the band should appear at the size 10kb after the vector is cut.
In addition, the sequence of the insert was confirmed in the sequencing result, but the sequence of the vector primer is not binding and the sequence cannot be confirmed.
I would like to ask your opinion on this.
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