Which bands for pET in agarosegel ? - (Feb/13/2007 )
Hi, can somebody tell which bands are normally seen on agarosegel when running uncut pET30a+ vector (Novagen) ?
I got the vector from another lab and transformed (the empty vector) into Top10 to be able to propagate it myself. I prepared a miniprep and also did a digest, now I got 1 band (~10kb) for uncut vector and 1 band (~6,5kb) for digested vector (regular size of vector is 5,4kb). Did anyone also propagate this vector themselve and experienced any problems / can the host strain have any influence ?
-pET-
QUOTE (pET @ Feb 13 2007, 11:17 AM)
Hi, can somebody tell which bands are normally seen on agarosegel when running uncut pET30a+ vector (Novagen) ?
I got the vector from another lab and transformed (the empty vector) into Top10 to be able to propagate it myself. I prepared a miniprep and also did a digest, now I got 1 band (~10kb) for uncut vector and 1 band (~6,5kb) for digested vector (regular size of vector is 5,4kb). Did anyone also propagate this vector themselve and experienced any problems / can the host strain have any influence ?
I got the vector from another lab and transformed (the empty vector) into Top10 to be able to propagate it myself. I prepared a miniprep and also did a digest, now I got 1 band (~10kb) for uncut vector and 1 band (~6,5kb) for digested vector (regular size of vector is 5,4kb). Did anyone also propagate this vector themselve and experienced any problems / can the host strain have any influence ?
the size of digested vector indicates that the vector as an insert. maybe the other lab gave you a vector with an insert that can still be used if you know the restriction sites that they have used.
the salts of the restriction buffer can alter the migration. but 1kb is a huge difference.
if this isnt the case, check the marker. count all the bands and see if you are missing one of them.
-nori_nediam-