What volume of vector and insert should be taken (in microliters) for ligation, if both the insert and vector amount is same in nanograms after gel extraction (say both are 12ng/μl)?
Michael J. Benedik
The mass of the vector and insert are not the relevant parameters, what is relevant is the molar concentration. So if you have similar mass of plasmid that is 5kb and insert that is 0.5 kb then your molar ration of insert to plasmid would be 10:1 when the optimal is more like 2:1 or 3:1.
But with regards to volume, in most cases the more concentrated the better. If you have too little DNA then you may not get good inter-molecular ligation.
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Papers
More Abdullah Khan's questions See All
Similar questions and discussions