excessive bps to NdeI

[Chris LaRosa]

zhongyi at sunm.shcnc.ac.cn wrote:

> Hello
>    if I want to improve the cleavable efficiency of primer with NdeI recognition site,how many excessive bps should be added to side of the NdeI recognition sequence?
> T

As a rule of thumb I always was told 1 extra is a fatal mistake... Add at least 3.  I usually add 5 extra base pairs.  See the back of the NEB catalog for charts on the
efficiency of cutting for specific enzymes.  I suggest you consult tech support from the enzyme manufacturer also.