Question

What would happen if:

1. I plugged the leads for the DNA gel in the wrong way? I was going too fast and I plugged the positive lead into the negative and the negative lead into the positive on the power supply?
2. I forgot one of my primers in the PCR tube? Would the PCR reaction work?
3. When I was extracting plasmid, I added the P3 buffer before adding the P2 buffer? Would the reaction still work?
4. After eluting the mtDNA, my partner and I mixed our samples together. We sent that mixed sample to be sequenced. Do you think we would get a sequence back?

Answer 1

For the first case, there will be no flow of current and therefore no separation of DNA fragments. This is because you have entered the wrong plugs because of which the current will not flow between the electrodes.

In the second case, PCR reaction will not work. It is because primer provide free hydroxyl group to taq polymerase to which it add deoxyribonucleotide and polymerises the daughter DNA molecule. So if primer is absent, hydroxyl group will also be absent and taq polymerase will not be able to work.

In the third care, you will still be able to extract plasmid but the quantity will be less. And it can also happen that there will be no effect on the quantity.

Mitochondrial DNA is highly conserved across individuals and between different species as well. So if you have given mixed sample for Sequencing, then first case will be that there will be no difference because of the conserved mitochondrial DNA is equal or there will be very less errors.

Please give a good rating.