I am trying to clone a 300 bp gene into pTYB2 vector (7.4kb)for later
protein purification method by IMPACT-CN. NdeI (producing sticky end) and
SmaI (blunt end) restriction sites were constructed into primers
corresponding to the N- and C- termini of the gene and incorporated by PCR
amplification. The PCR product and the vector were both double digested with
SmaI and NdeI respectively. After gel purification, the digested insert (
gene) and the vector were used to perform ligation with T4 DNA li
