Question

72)PCR primers, Primer-F and Primer-R were supplied with the following specifications from Sigma Inc., Specifications Primer F Primer R ug/OD 30.3 35.6 ug 262 337 ul for 100uM 421 565 Each primer was first dissolved in 1000 ul of TE buffer and then 10x times diluted. 2ul of each primer was then added to a PCR reaction mix (Total reaction volume = 25ul). What is the final concentration of primer “F "and “R” in the PCR reaction mix? A. 52.4 and 67.4 ng B. 52.4 and 67.4 49 C. 26.2 and 33.7 ng D. 52.4 and 67.4 pg​

Answer 1

Explanation:

. Centrifuge the primer tubes, 8000 rpm for 30 seconds to get all the lyophilised DNA to the bottom of the tube.

2. Dissolving:

For primer sequence

Primer 20.91 nmol

add 20.91x 10 microL, equals to 209 microL of sterile ultrapure water

(it makes 100 micromolar Stock soln or 100 pmols/ microlitre- 1)

3. Allow to sit for 2mins, keep in the water bath at 60 degree centigrade for 10

minutes.

4. Again centrifuge 8000 rpm for 30 seconds.

5. Take 10microlitre of this (above 100 pmols/ microlitre primer- 1) and add to it 90 microlitre of ultrapure water. Gives you the working solution 10 micromolar Stock

soln or 10 pmols/ microlitre)

working solution--------Primer 1A.

(The oligonucleotide dissolved in water is stable for at least 6 months at -20°C)

6. From 1A use for PCR mix.

I hope it is simple enough.

Make me as brainliest... All the best