I performed a ligation earlier that will be used in a transformation tomorrow. However, I made an error in reading the protocol and mistakenly added twice as much 10X T4 Ligation Buffer in mixing with my plasmid, diluted duplex oligo, dH20, and T4 ligase. Specifically, the protocol asked for 50ng plasmid, 1ul oligo, 1ul ligase, 1ul buffer, and dH20 for a total of 10ul system.
However, instead of 1ul I added 2ul of 10X T4 ligation buffer. I understand that the main purpose of the buffer is to provide ATP, but I'm unsure if twice as much would actually result in any noticeable negative effects.
Unfortunately, I have no more plasmid to be able to redo the ligation. I still intend to perform the transformation tomorrow, but would like to know maybe what I can expect or if it's even that good idea to continue. I'm probably overreacting, but this particular experiment is pretty important.