Posted 19 July 2002 - 06:36 AM
Posted 23 July 2002 - 07:14 PM
dephos enzyme and buffer from Lifetech (GibcoBRL)
Posted 04 August 2002 - 05:19 AM
2- you could try by PCR. amplify your insert with primers having enzymatic restriction sites to obtain cohesive ends compatible with your vector.
3- your insert is very big. It can be a problem. You can try to do ligation in two times. Cut you insert in two pieces (1,6Kb) and insert one after each other.