DNA extraction

[j7235]

I have followed the kit instruction, however, after I added the elution buffer to both of the 2 samples (from the same stem of the plant) for DNA extraction, I find out one of the AC column contains more liquid than the other one, so I pipette the excessive liquid from one AC column (for one sample) to another one (for another sample). It seems to be not a good idea, but does it mean that the two samples cannot be used?

In another DNA extraction, I added the elution buffer not to the centre of the AC column (suggested by the instruction in the kit), does it mean that I better discard these samples?

I only tried DNA extraction for three times... so... 

Wish someone could help... Thanks!

[CharonY]

There are dozens of different protocols so it is a bit difficult to talk about specifics. But generally speaking, merging several samples is fine as long as you do not overload the column and provided that you can actually mix the samples (obviously if they are supposed to be different samples, do not mix them). But I would first look whether  you got too much material or debris in your sample that may clog the column. 

With regards to the elution buffer, think about why they would suggest pipetting it there. And what are the worst case scenarios if you do not? 

[j7235]

On 9/4/2019 at 5:16 AM, CharonY said:

There are dozens of different protocols so it is a bit difficult to talk about specifics. But generally speaking, merging several samples is fine as long as you do not overload the column and provided that you can actually mix the samples (obviously if they are supposed to be different samples, do not mix them). But I would first look whether  you got too much material or debris in your sample that may clog the column. 

With regards to the elution buffer, think about why they would suggest pipetting it there. And what are the worst case scenarios if you do not? 

Thanks for your response and I figure out the elution buffer is added to the centre of column only for better elution of the DNA from the membrane for centrifuge and adding it to the side may cause some of the DNA not effectively eluted from the membrane but DNA can still be extracted as long as some elution buffer is added to the membrane for centrifuge. No big deal.