Dear researchers please help
Im actual;y working on cold active lipases. Currently trying to look for novel lipases. Here is what i did:
I did intial digest with Sau3A1 with my genomic DNA with different concentration and mange to get a band of 10kb to 2 kb in which the lipase genes are believed to be in.
I ligated my product into puc 19 (vector) and host of DH5 alpha. I seem to get good no of colonies on the plate however when I test with RE digest (double digest) i just get a smearing band. Im using Eco R1 with Hind111 buffer tango
Please help!!! Do i change the plasmid, or the RE?