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 μl 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.
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 μl 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.
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Control of Microbial Growth
It can be defined as the process to inhibit or prevent the growth of the population of microorganisms. It usually involves the use of physical and chemical agents to the growth of microorganisms. It is very important to control the growth of microorganisms, especially in the pharmaceutical and biotechnology industries, academic research, medical field, and food industry.
Sterilization
It is a method for the destruction of all forms of vegetative growth and endospores from a medium. In other words, sterilization can be defined as a method of killing every microbial organism such as bacteria, fungi, etc. present in an object or a medium. There are different strategies present to sterilize an object, like physical methods and chemical methods of microbial control.
Question
Caculate the concentration of DNA of each tube (1-8) at the end, including the unkown.
The DNA concentration is 1.5
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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