i am getting no colonies after ligation of 1.2kb insert in to 9.4kb vector cut with bam H I and Sph-I iam useing insert : vector ratio 8:1.
ligation reaction before ligation shows two band one of 9kb and one of 1kb
immidiate after ligation i got three bands one of 9kb one of 3kb and one of 1 kb.
after overnight ligation reaction i got two more very faint bands of 5kb ad 8kb.
my transformation efficaccy for control plasmid was 4.7 X 10 raised to the power 3.
please tell me whether my ligation worked or i have onley get nospecific ligated product ?
if the ligation occured then is it due to low transformation efficaccy?