For the reverse transcription reaction, the components below are assembled to ma

Question

For the reverse transcription reaction, the components below are assembled to make a 2X reverse transcription reaction mix as shown in the table (for ONE reaction). To this mix, 10 mu l of RNA is added. Calculate the following (and show your work): a. What "X" concentration is the reaction solution after the addition of 10 mu l of RNA? b. In lab, you normalized all of your RNA samples to a single concentration. Using this concentration, what is the total amount of RNA that will be added to the cDNA reactions next week when you perform cDNA synthesis? c. What is the concentration of RNA in the 20 mu l reaction? d. Assuming 100% complete conversion of RNA to cDNA, what is the concentration of cDNA when the reaction is complete?

Explanation / Answer

5.

A. after adding 10 microlittre RNA the Reaction mixture will be 1X.

B. If all the samples are normalized then we should use the same amount of RNA next week also.

C. We can not calculate the Final concentration without knowing the initial or stock concentration which is not mentioned in the question.

D. Same is true for this question also.

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