why the inserted fragment can not be cutted ..... - the inserted fragment can not cutted by the enzymes used (Nov/22/2005 )
Hi, I have a strange question which is always disturbing me very much.
I clone a PCR fragment in to the vector by using ECORI and BGL II. comparing the colony number on the plate of the vector only control and that of vector + insert , It seems the cloning is perfect.
also, using PCR , we got the insert band. but why some clone I could not digested by ECORI AND BGL II. the sites corresponding to these enzymes are less like be methlated , How could ? actually, the thing happened to me several times before.
dose any one can explain the reason?
Thanks
-Flox-
How big is the insert and how big is the vector? Do you have an image of a gel showing the digestion to upload?
QUOTE (Flox @ Nov 23 2005, 07:10 AM)
Hi, I have a strange question which is always disturbing me very much.
I clone a PCR fragment in to the vector by using ECORI and BGL II. comparing the colony number on the plate of the vector only control and that of vector + insert , It seems the cloning is perfect.
also, using PCR , we got the insert band. but why some clone I could not digested by ECORI AND BGL II. the sites corresponding to these enzymes are less like be methlated , How could ? actually, the thing happened to me several times before.
dose any one can explain the reason?
Thanks
I clone a PCR fragment in to the vector by using ECORI and BGL II. comparing the colony number on the plate of the vector only control and that of vector + insert , It seems the cloning is perfect.
also, using PCR , we got the insert band. but why some clone I could not digested by ECORI AND BGL II. the sites corresponding to these enzymes are less like be methlated , How could ? actually, the thing happened to me several times before.
dose any one can explain the reason?
Thanks
-ML1975-