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C. Estimating DNA concentration - creating a 2-fold dilution series. Can you estimate how much DNA is an unknown sample? 1. Label an 8-tube strip of PCR tubes Label the first seven tubes 1-7. Label the final tube "U" for unknown.

C. Estimating DNA concentration - creating a 2-fold dilution series. Can you estimate how much DNA is an unknown sample? 1. Label an 8-tube strip of PCR tubes Label the first seven tubes 1-7. Label the final tube "U" for unknown.

Biology: The Unity and Diversity of Life (MindTap Course List)
Biology: The Unity and Diversity of Life (MindTap Course List)
15th Edition
ISBN:9781337408332
Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Chapter15: Studying And Manipulating Genomes
Section: Chapter Questions
Problem 16SQ: Match the method with the appropriate enzyme. _____ PCR a. Taq polymerase _____ cutting DNA b. DNA...
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Caculate the concentration of DNA of each tube (1-8) at the end, including the unkown. 

C. Estimating DNA concentration - creating a 2-fold dilution series. Can you estimate how much DNA is an unknown sample? 1. Label an 8-tube strip of PCR tubes • Label the first seven tubes 1-7. Label the final tube "U" for unknown. 2. To tube 1, add 10 µl of 50:50 AT:GC DNA (tube B). 3. Add 5 µl of Buffer to tubes 2-7. 4. Remove 5 µl from tube 1 and add it to tube 2. • Pipette up and down gently three times to mix. 5. Remove 5 ul from tube 2 and add it to tube 3. • Pipette up and down gently three times to mix. 6. Continue diluting samples in the 8-tube strip. Remove 5 µl of sample from tube 3 and add it to tube 4. Mix gently. • Remove 5 ul of sample from tube 4 and add it to tube 5. Mix gently. Remove 5 ul of sample from tube 5 and add it to tube 6. Mix gently. 7. Remove 5 µl of sample from tube 6 and discard. Tubes 1-6 should all have 5 µl of sample in each tube. • Tubes 1-6 now have a 2-fold serial dilution series. • Tube 7 should have no DNA and will serve as a blank control. 8. Add 5 µl of the "Unknown DNA Concentration" to tube U. • The teacher will have "Unknown DNA Concentration" at the front of the room. 9. Add 10 µl of Dye to all 8 tubes. 10. Add 10 µl of Buffer to all 8 tubes. 11. Cap the 8-strip of PCR tubes and place in P51TM or other blue light illuminator. • Darken the room, or use a light blocking hood to better view the samples. 12. Estimate the concentration of DNA in the unknown from your dilution series. Concentration of 50:50 AT:GC DNA before adding to tube 1 was 1.5 µM (micromolar). • Brightness of the unknown sample is likely to not match any one tube in the dilution series exactly. Use a best approximation.
Transcribed Image Text:C. Estimating DNA concentration - creating a 2-fold dilution series. Can you estimate how much DNA is an unknown sample? 1. Label an 8-tube strip of PCR tubes • Label the first seven tubes 1-7. Label the final tube "U" for unknown. 2. To tube 1, add 10 µl of 50:50 AT:GC DNA (tube B). 3. Add 5 µl of Buffer to tubes 2-7. 4. Remove 5 µl from tube 1 and add it to tube 2. • Pipette up and down gently three times to mix. 5. Remove 5 ul from tube 2 and add it to tube 3. • Pipette up and down gently three times to mix. 6. Continue diluting samples in the 8-tube strip. Remove 5 µl of sample from tube 3 and add it to tube 4. Mix gently. • Remove 5 ul of sample from tube 4 and add it to tube 5. Mix gently. Remove 5 ul of sample from tube 5 and add it to tube 6. Mix gently. 7. Remove 5 µl of sample from tube 6 and discard. Tubes 1-6 should all have 5 µl of sample in each tube. • Tubes 1-6 now have a 2-fold serial dilution series. • Tube 7 should have no DNA and will serve as a blank control. 8. Add 5 µl of the "Unknown DNA Concentration" to tube U. • The teacher will have "Unknown DNA Concentration" at the front of the room. 9. Add 10 µl of Dye to all 8 tubes. 10. Add 10 µl of Buffer to all 8 tubes. 11. Cap the 8-strip of PCR tubes and place in P51TM or other blue light illuminator. • Darken the room, or use a light blocking hood to better view the samples. 12. Estimate the concentration of DNA in the unknown from your dilution series. Concentration of 50:50 AT:GC DNA before adding to tube 1 was 1.5 µM (micromolar). • Brightness of the unknown sample is likely to not match any one tube in the dilution series exactly. Use a best approximation.
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