Restriction digestion of clones give undesired product
Posted 19 September 2011 - 02:45 AM
ideally i should have got inserts of 2.6 kb in all of them . but i got it only in one of them . and in the other i got bands at around 1.5 KB .
my question is what is wrong ?
how can T vectors give transformed colonies if they r not ligated ?
if they have ligated it should b the product i have gel purified then Wat is the band around 1.5 ?
has there been a wrong insert ? but how is it possible i did gel purification and reconfirmed the band by running it on gel and getting desired band .
Posted 19 September 2011 - 05:08 PM
What controls did you run for your ligations?
Posted 21 September 2011 - 02:04 AM
none of Kpn 1 and Bsu 361 are reported of star activity- i checked that ..
i did not run any control for ligation .. why would that be necessary?
Posted 21 September 2011 - 05:02 PM
The vector can self ligate - this is quite common with complementary ends.
Perhaps your PCR product wasn't what you thought it was. Try sequencing the vectors you have and see if there is a problem or not.