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You need to perform PCR to amplify a gene for this assignment. You will create a PCR solution with 25 µL total volume. The recipe for the PCR solution is below: PCR recipe 10X PCR master mix stock DNA (125 ng total DNA) Forward primer (400 nM final concentration) Reverse primer (400 nM final concentration) Part 1 Your stock DNA concentration is 1.25 µg/μL. You want to dilute it to 50 ng/μL as a working solution that you then will use in your PCR reaction. How would you make the 50 ng/µL DNA solution using only one dilution step? Indicate the dilution factor in your answer. Show your work. Note, you must use units and explain how you would make this in the lab (include the diluent). You must make a large enough volume so that you have enough to use in your final PCR solution.

You need to perform PCR to amplify a gene for this assignment. You will create a PCR solution with 25 µL total volume. The recipe for the PCR solution is below: PCR recipe 10X PCR master mix stock DNA (125 ng total DNA) Forward primer (400 nM final concentration) Reverse primer (400 nM final concentration) Part 1 Your stock DNA concentration is 1.25 µg/μL. You want to dilute it to 50 ng/μL as a working solution that you then will use in your PCR reaction. How would you make the 50 ng/µL DNA solution using only one dilution step? Indicate the dilution factor in your answer. Show your work. Note, you must use units and explain how you would make this in the lab (include the diluent). You must make a large enough volume so that you have enough to use in your final PCR solution.

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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Please help me with question number one. Thank you so much!

**PCR Amplification Assignment** For this assignment, you are tasked with performing PCR to amplify a gene. You will create a PCR solution with a total volume of 25 µL. Below is the recipe for the PCR solution: **PCR Recipe:** - 10X PCR master mix stock - DNA (125 ng total DNA) - Forward primer (400 nM final concentration) - Reverse primer (400 nM final concentration) **Part 1** Your stock DNA concentration is 1.25 µg/µL. You need to dilute it to 50 ng/µL as a working solution for your PCR reaction. - **Task:** Make the 50 ng/µL DNA solution using only one dilution step. Indicate the dilution factor and show your work. *Note: Use proper units and explain the process, including the diluent. Ensure the volume is sufficient for the final PCR solution.* **Part 2** The stock primer concentrations are both 100 µM. Dilute them to 5 µM as a working solution for your PCR reaction. - **Task:** Make the 5 µM primer solution using only one dilution step. Indicate the dilution factor and show your work. *Note: Use proper units and explain the process, including the diluent. Ensure the volume is sufficient for the final PCR solution.* **Part 3** Using the diluted DNA and primer solutions from Parts 1 and 2, calculate the recipe for your 25 µL PCR reaction. Use the components above (see PCR recipe) to solve this. Show your work.
Transcribed Image Text:**PCR Amplification Assignment** For this assignment, you are tasked with performing PCR to amplify a gene. You will create a PCR solution with a total volume of 25 µL. Below is the recipe for the PCR solution: **PCR Recipe:** - 10X PCR master mix stock - DNA (125 ng total DNA) - Forward primer (400 nM final concentration) - Reverse primer (400 nM final concentration) **Part 1** Your stock DNA concentration is 1.25 µg/µL. You need to dilute it to 50 ng/µL as a working solution for your PCR reaction. - **Task:** Make the 50 ng/µL DNA solution using only one dilution step. Indicate the dilution factor and show your work. *Note: Use proper units and explain the process, including the diluent. Ensure the volume is sufficient for the final PCR solution.* **Part 2** The stock primer concentrations are both 100 µM. Dilute them to 5 µM as a working solution for your PCR reaction. - **Task:** Make the 5 µM primer solution using only one dilution step. Indicate the dilution factor and show your work. *Note: Use proper units and explain the process, including the diluent. Ensure the volume is sufficient for the final PCR solution.* **Part 3** Using the diluted DNA and primer solutions from Parts 1 and 2, calculate the recipe for your 25 µL PCR reaction. Use the components above (see PCR recipe) to solve this. Show your work.
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