Question

A student does a PCR to make copies of the HDR template transformation. They column-clean the PCR, eluting in 20 ul of warm elution buffer. They use 2 ul of the 20 ul stock to nanodrop the sample, and determine the concentration is 60 ng/ul. How many nanograms of HDR template does the student have left to use for future work (e.g. yeast transformations)

Answer 1

the student will have 1080 nanograms of HDR template left to use for future work

Explanation:

Given that:

the total volume of DNA which is eluted after the PCR clean up = 20ul

PCR clean up is a procedure by which the PCR product is gotten rid of DNA polymerase enzyme, excess primers, excess dNTPs so that these does not interfere in further experiments such as transformation.

using 2 ul of the 20 ul stock to nanodrop the sample

Concentration of the DNA after the PCR clean up = 60ng/ul

No, Since 2ul is used in the nanodrop, thus total volume left for further experiment will be = 20 - 2 = 18ul

Now, the total amount of the HDR template the student have left to use for future work will be;

⇒ 18ul × 60 ng/ul

= 1080 ng

Therefore, the student will have 1080 nanograms of HDR template left to use for future work