You have a 5 mM stock of MgCl2 and you want to prepare a 30 ul...
You have a 5 mM stock of MgCl2 and you want to prepare a 30 ul mixture of PCR reagents that has a final concentration of 100 uM of MgCl2. How much of the stock MgCl2 would you need to add to the PCR mixture?
Homework Answers
According to the formula:
M1V1 = M2V2,
5 * 10-3 * V1 = 100uM * 30
V1 = 100*30/5 * 10-3 = 0.6 ul.
Thus a total of 0.6ul of stock solution can be used for the working reaction to make a solution of 30ul. However, this volume cannot be pipetted out so the stock solution can be diluted at least 10 times to get a working diluted solution, and then a volume of 6ul can be pipetted for the working reaction.
2. You are performing PCR under the following conditions: 1. Each PCR reaction contains 50 pl...
2. You are performing PCR under the following conditions: 1. Each PCR reaction contains 50 pl final volume II. Each PCR reaction contains: 1x PCR buffer, 1.5 mM MgCl2, 0.2 mM dNTP mix, 0.2 MM sense primer, 0.2 uM anti-sense primer, 1.5 units of Taq polymerase and 2 ul of DNA template. [all concentrations are final] III. Stock solutions of the following reagents are available: 10x PCR buffer, 25 mm MgCl2, 10 mM dNTP mix, 10 M of sense primer,...
Question 23: &n
Question 23: (10 Marks) a) Given the stock concentration and desired final amount, or desired final concentration, of the following PCR components, calculate the volume of each you would add to a 25 μl PCR reaction mixture. b) You want to analyse the resultant PCR product by agarose electrophoresis. Calculate how much agarose you need to use to make up 50 ml of 1.5 % gel. How much of 5x concentrated loading buffer will you add to load the...
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Molecular Key Calculation - PLEASE NEED
HELP TO CALCULATE IT!
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In each digestion tube you will be adding 10 ul DNA and 40 ul of
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You are given 10 x Not1 buffer, 10 ug/ul acetylated BSA, Not1
restiction enzyme 10 units/ul and sterile water.
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Nucleic Acids 33 boslari, no NO E Let's Make a PCR Reaction Reaction Mixture: You have stock DNA of 100 ng/uL that you wish to amplify. You have stocks of 10X PCR Buffer, 10 uM of the forward and reverse primers, 25 mM MgCl2, 50 mM dNTPs , and Tag that is 10 Units/UL. Create the Recipe for a 20 PL total PCR that uses 50 ng of DNA, 2.5 mM primers, 5 mM MgCl2, 5 mM dNTPs,...
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Prepare a fragment of DNA to be cloned by PCR by preparing the
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please hlep me answer those three questions asap.
Please ignore question 7.
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B 2 ml 500 ul 1 1.25 ml 500 ul 1 ml Reagent HO 300 mM...
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Question 6 Using stock solutions to make up a solution: from final concentration to volume You need to digest a sample of DNA with a restriction enzyme. You will do this reaction later in the semester. The buffer for this reaction has a final concentration of 40 mM Tris, pH 7.5, 10 mM MgCl2 and 50 mM KCl. You have the following stock solutions: 0.5 M Tris, pH 7.5, 1 M MgCl2 and 2 M KCl. How would you make...
2. You are performing PCR under the following conditions: 1. Each PCR reaction contains 50 pl final volume II. Each PCR reaction contains: 1x PCR buffer, 1.5 mM MgCl2, 0.2 mM dNTP mix, 0.2 MM sense primer, 0.2 uM anti-sense primer, 1.5 units of Taq polymerase and 2 ul of DNA template. [all concentrations are final] III. Stock solutions of the following reagents are available: 10x PCR buffer, 25 mm MgCl2, 10 mM dNTP mix, 10 M of sense primer,...
Molecular Key Calculation - PLEASE NEED
HELP TO CALCULATE IT!
You are setting up you own Master Mix for doing a Not1
restriction digest.
In each digestion tube you will be adding 10 ul DNA and 40 ul of
your Master mix.
You are given 10 x Not1 buffer, 10 ug/ul acetylated BSA, Not1
restiction enzyme 10 units/ul and sterile water.
How much of each of the 4 reagents would you use to make 500 ul of...
biochemistry question
Nucleic Acids 33 boslari, no NO E Let's Make a PCR Reaction Reaction Mixture: You have stock DNA of 100 ng/uL that you wish to amplify. You have stocks of 10X PCR Buffer, 10 uM of the forward and reverse primers, 25 mM MgCl2, 50 mM dNTPs , and Tag that is 10 Units/UL. Create the Recipe for a 20 PL total PCR that uses 50 ng of DNA, 2.5 mM primers, 5 mM MgCl2, 5 mM dNTPs,...
Prepare a fragment of DNA to be cloned by PCR by preparing the
oligonucleotides received through dilutions. The DNA to be used as
a template is in a 50 ng/ul solution. The protocol is as
follows:
VOLUME TO ADD FINAL CONCENTRATION 0.5 UM 0.5 M REAGENT 10 PM Forward Primer 10 PM Reverse Primer Thermoestable pol Master mix 2x Template DNA water 1x 100 ng.. Total volume 25 ul Information about the primers: Forward primer: 29.3 nmoles - 220 ug...
please hlep me answer those three questions asap.
Please ignore question 7.
Question 7 Working with buffers You need 100 ml of 120 mM phosphate buffer and you have two stock solutions from which you can make the buffer; 0.6 M NaH2PO4 (the acid form) and 0.6 M Na2HP04 (the base form). You need to add 3 times more of the base form than the acid form to achieve the desired pH. How will you make this solution? Question 8...
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