i have problem for ligation for 2 months. 'plz help and suggest me.
now , i wanted to just chek whether the ligATION is working or not.
i wanted to ligate avian RSK1 into PKH3 vector .for this i cut both with BamH1 and Hpa1 enzyme .
i cut two bands in gel electrophoresis. i also ligated both cuts ( BANDS) of PKH3 as a positive control and AND ALSO pkh3 as vector and RSKN1 as insert.
i did transformation.
Unfortunately i did not get any colony.
i suffered this for 2 month .
i couldnot find which step is wrong and where is the proplem? i got frusted . Please suggest me