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Basic Clinical Laboratory Techniques 6E

Basic Clinical Laboratory Techniques 6E

6th Edition

ISBN:9781133893943

Author:ESTRIDGE

Publisher:ESTRIDGE

Chapter3: Basic Hemostasis

Section3.6: D-dimers

Problem 8RQ

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Show the calculation for the preparation of 0.01%,
0.001%, 0.0001%, 0.00001% standard DNA solutions
via serial dilution from a 0.1% stock solution.

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You have a 20 mg/ml of Ethidium bromide stock solution. You need to a final concentration of 2ug/ml into a Agarose solution to visualize DNA. What is the dilution factor? Give typing answer with explanation and conclusion
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A 10 μL-aliquot of a resuspended genomic DNA stock solution was obtained and further diluted by adding 990 μL TE buffer. The A260 of the resulting solution was 0.316. Determine its concentration in µg/µL.
You will use a Calf Thymus DNA standard stock solution (1 mg/ml) to prepare a standard curve. Calculate volume of stock solution in ul, needed to prepare 10 ug/ml dilution (final volume 1.0 ml).
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You have a very concentrated sample of DNA which you need to dilute for sequencing 1:6000 with a final volume of 10 μl. How would you do this in four steps, using a serial dilution? Do not pipet less than 1 ul and do not make more than 10 ul per dilution. Describe how to do each dilution using the format suggested below.
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Based on the quantitation data, please indicate how you would dilute the following samples. Fill in each blank with the appropriate information. Item 1: DNA concentration - 22.6 ng/uL Dilution factor - 1: Add uL DNA extract to uL buffer Final dilution concentration = ng/ul Item 4: DNA concentration - 3.75 ng/uL Dilution factor - 1: Add uL DNA extract to uL buffer Final dilution concentration = ng/ul
One OD unit of A260 is 50 ug/ml dsDNA. You have 2.75 ml genomic DNA (stock). You took 50 ul of the DNA sample and mixed it with 950 ul water and measured the A260 absorption. The diluted sample absorbed 0.14 units of A260. The DNA concentration of the stock is ug/ml, considering the dilution factor of (Note: ug microgram, ul- microliter). 70/10 140/20 280/50 O 560/100
In order to determine which dye (Ethidium bromide (10 µg/ml), EvaGreen (10 µg/ml) or SYBR Green (10 µg/ml) is the safest to use in a laboratory for DNA binding, first sterilise the spreader and forceps with 95% ethanol, then place 2-3 drops of bacterial culture on each plate and spread well with a sterile spreader and using the sterile forceps, place a sterile filter disk on the center of each plate. Next, Add a drop of sterile water to the filter paper on the control plate, a drop of ethidium bromide to another plate, a drop of EvaGreen to another plate, and a drop of SYBR green to another plate. The sterile water is used as a negative control. And then incubate the plates at 37°C for 48 hours. The results obtained are attached as a picture, after looking at the results obtained in the picture; use the Ames Assay to determine which of the provided dyes are safest?
To carry out sequencing, you need to include 600 ng of DNA and a total of 6.4 pmol of sequencing primer. If your concentration of DNA (determined by A260 reading on a Nanodrop Spectrophotometer) is 89.0 ng/μL, how many μL of this sample do you need for a total of 600 ng?

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