I have 122.7 ng of insert and 227 ng of vector both of which has to be ligated. I'm confused as to how much volume of both vector and gene of interest has to be considered before setting up the ligation reaction. Please help!
The idea is that regardless of size, each linear DNA molecule only has two ends, and thus it is end concentration that matters in the ligation reaction.
Hi
Better to calculate the number of moles of vector and insert per microlitre and then use it in the ligation reaction with a ration of vector to insert as 1:2 or 1:3.
For calculating the number of moles of either vector or insert you must know their sizes and their concentration. There are number of tools available to calculate, moles of a fragment based on their size and concentration.
Best luck.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology