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To collect DNA you need to lyse the cells with a lysis buffer. To make this you first prepare stock solutions of NaCl (4M), Tris (1M, pH 8), 10% SDS (w/v) and Na2 EDTA (dehydrate)(0.5M). How would you make 100 mL of each stock solution? Note: Look up molecular weights.

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Beverley Smith
Beverley SmithLv2
29 Sep 2019

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Related questions

You have just begun working in a research laboratory. Your advisor asks you to make a series of stock solutions. He also explains that you may use a pH meter to adjust the pH. Calculate how much of the solid reagent you would add to make the following:

500 mL 1 M Tris, pH 8.0 (MW = 121.4 g/mole)

1.0 L 5 M NaCl (MW = 58.44 g/mole)

10 mL 100 mg/mL Ampicillin (MW = 371.4 g/mole)

500 mL 0.5 M EDTA (Ethylene Diamine Tetra-acetic Acid; MW = 292.2 g/mol)

200 mL 1 M MgCl2 (Note: MgCl2 is sold by the chemical company as co- crystallized with H20. Thus, the MW of MgCl2 6H20 is 203.30 g/mole)

For your first experiment, you need to make a solution that is commonly called TE and stands for Tris-EDTA. It is comprised of 10 mM Tris, 1 mM EDTA, pH 8.0. Calculate and describe how you would make 100 ml of TE using the stock solutions that you have already made above in Question 1.

Your advisor asks you to make two concentrated stock solutions for the whole laboratory to use. He explains that it is common to make a concentrated solution to dilute for use. Thus, a 10x solution refers to one that is 10-fold concentrated. Calculate how you would make 1.0 L of each of the following:

A. 10x TBE (0.9 M Tris, 0.9 M Boric Acid, 20 mM EDTA) Tris MW = 121.4 g/mole

Boric Acid MW = 61.84 g/mole EDTA MW = 292.2 g/mole

B. 50x TAE (2 M Tris, 2 M Acetic acid, 0.5 M EDTA) Tris MW = 121.4 g/mole

Concentrated Acetic acid (glacial) comes as a solution and is 17.4 M EDTA MW = 292.2 g/mole

Your polyacrylamide gel electrophoresis experiment requires 1.0 L of 0.5x TBE. Calculate and describe how you would make this solution using your stock solutions

Assignment: For each question, describe how you would make the solution using the stocks or reagents listed below. On our lab prep shelves, we have the following solutions and powders:

Sodium citrate (FW = 43.7 g/mol)

SDS powder (FW = 288.38 g/mol)

0.5 M EDTA IPTG powder (FW = 238.3 g/mol)

20 mg/mL X-gal solution

1000X ONPG

Premade LB

Premade LB agar

0.5 mM Colchicine

50X gel running buffer

Agarose powder

95% ethanol

100 mg/mL Ampicillin

10X BamHI restriction enzyme digestion buffer

milliQ water dimethylformamide (DMF)

Example 1. (DNA isolation lab) How would you make 100mL of 0.15M Sodium Citrate, 5% SDS, 0.001M EDTA solution (one solution that contains all of these reagents)? Answers 0.65 g Sodium citrate 5 g SDS 0.2 mL of 0.5 M EDTA Add 75 ml of water and mix well. Then add water up to 100 ml to complete.

Example 2. (B-gal) How would you make a 10 fold dilution of 20 mg/ml X-gal? X-gal is not soluble or stable in water, but it is in dimethylformamide. What would the concentration of the diluted solution be? Answer Add 1 ml of the stock solution to 9 ml of DMF. 2 mg/ml

Problem for you

1. How would you make 500 mL LB = 40ug/mL X-Gal? ( the LB is made, so you just need to figure out how much X-Gal to add)

How would you make 500mL LB = 0.1mM IPTG? You cannot add solid IPTG to media. Instead use a sterile 1M stock of IPTG

How would you make 500mL LB + 40ug/mL X-gal + 0.1mM IPTG?

Please show all work that got you to your solution

thank you!

A)Describe how you would prepare a 1X TBE electrophoresis bufferusing the 10X TBE as a stock solution assuming you need a finalvolume of 800 ml.

B) Assume that a 0.5% (w/v) stock solution is available in thelab that you will use to make up three different 5 ml workingsolutions. The concentration of each of the working solutions is asfollows: 0.25%, 0.05%, and 0.001%. Calculate the volume of stocksolution and water that will be used for each dilution. Indicatewhich micropipette you would use to make each working solution.

C) Assume that you need to mix a 1 mL stock solution of 10Xrestriction buffer with the following final concentration: 600 mMTris, 100 mM MgCl, 70 mM DTT. Further assume that you have thefollowing solutions available to you: 2 M Tris, 1M MgCl, and 1.5 MDTT. Describe the preparation of the 10X restriction buffer showingall calculations. Describe how you would dilute this 10Xrestriction buffer to 2X with a volume of 50 microliters. Rememberthese volumes are much too small for volumetric flasks.

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