H. You are setting up four PCR reactions using nested GAPDH primers. The reactions are summarized in the table below: Ne

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H You Are Setting Up Four Pcr Reactions Using Nested Gapdh Primers The Reactions Are Summarized In The Table Below Ne 1 (171.58 KiB) Viewed 156 times

H. You are setting up four PCR reactions using nested GAPDH primers. The reactions are summarized in the table below: Negative Control Tube 1 (ul) 10 Sample # 1 Tube 3 (ul) 10 Positive Control Tube 2 (ul) 10 5 4 Specimen #2 Tube 4 (ul) 10 0 2x Master Mix DNA Template H20 Taq Total 9 9 0 9 0 1 1 1 1 20 20 20 20 a. What is Tube 1 controlling for? (What would it mean if Tube 1 produced a PCR product?) b. What is Tube 2 controlling for? (What would it suggest if Tube 2 did not produce a PCR product?) c. You made the 2X Master mix before setting up your 4 reaction tubes. To be on the safe side, you make up enough 2X MM for 5 reactions. The master mix consists of 10X PCR buffer, forward and reverse primers, sterile water and (What is missing from the list?) How much 10x Buffer did you add to your 2x MM? (Hint: How much 10x buffer needs to go into each 20 ul reaction to make it a 1x solution?] d. Commercially available Master Mix usually comes in 2X concentration without the primers added. Why? (Why don't they just sell it with everything already added?)