Question

g You are performing DNA sequencing reactions as part of your job as a employee at GeneTech, Inc. In your current project, you are attempting to sequence a 700 bp DNA fragment sent by a customer. After the reaction has begun, you realize that due to a decimal error, you accidentally added the fluorescently labeled di-deoxyribonucleoside triphosphates (ddATP, ddGTP, ddCTP, ddTTP) at ten times higher than the desired concentration. What do you expect will be the consequence of this mistake?

Answer 1

The consequence of this error will be the waste of material, since several nucleotides were placed in the reaction and will not be used.

Step-by-step explanation:

There are several DNA sequencing protocols and although we don't know which one the GeneTech, Inc. employee is using, we can say that the fluorescence-labeled di-deoxyribonucleoside triphosphates will be used to make countless copies of the DNA particle that he wants to sequence.

As we know, DNA replication separates the DNA strands and uses each strand to make copies of new DNA molecules. These copies are made with the DNA polymerase enzyme by adding deoxyribonucleosides to the new tapes. As the enzyme is highly regulated and has a DNA strand as a template for the new strands, it will not add nucleotides beyond what is necessary.

Therefore, by placing excess di-deoxyribonucleoside triphosphates in the reaction, the consequence will be just the waste of product.