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10. Complete this Master Mix table for 3 DNA samples, a positive control,...
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,...
How to solve for question#6? 5 5. Fill in the table below to set up the...
How to solve for question#6?
5 5. Fill in the table below to set up the reactions for a single and then a set of PCR reactions: Concentration in Volume in 1 Master Mix for four Reagent Stock concentration 10X 25 mM one reaction 1X 2 mM reaction 25 jl reactions Buffer MgClz dNTPs Primer mix DNA template Taq polymerase water Total volume 2.SA 2 al 2.S A Pe 100 nM-0.IA 2 ng/HI 1 unit VS 10 μΜ 0.25 l...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in th...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
solve for gel preparation and PCR mastermix calculations with steps for understanding TBE Buffer Calculations Determine...
solve for gel preparation and PCR mastermix calculations with
steps for understanding
TBE Buffer Calculations Determine the mass of the following reagents for a 10X stock 700mM of Tris Base (157g/mol) 887mM of Boric Acid (62g/mol) 25.7mM of EDTA (292g/mol) Dissolve in 750ml of DIH.O and bring to volume (IL) Calculate the dilution of your 10x stock for a 1X working stock. Remember you only need IL of working stock for a single experiment. Gel Preparation Calculations You need to...
2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations...
2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations for calculating your cocktail. Fill in the chart below. A. Fill in the chart below with the volumes that would be added to each tube. Note the final volume of the reaction. Show your calculations for full credit. REAGENT Final Concentration (50 ul Reaction) 1x Test Reaction (+) Control Reaction 10x Reaction Buffer ML dNTPs (15 mm) 200 UM 0.2 UM ul 0.2 um...
1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteina...
1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteinase K; guanidine HCI; SDS 2. Why is the ratio of the OD at 260 and 280 nm used to estimate DNA purity? 3. In one paragraph, summarize basic principles of PCR technique in your own words. List all the reagents you will need to perform a PCR experiment. Does this method tell you what genetic modifications were made? If yes, describe how you can...
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,...
How to solve for question#6?
5 5. Fill in the table below to set up the reactions for a single and then a set of PCR reactions: Concentration in Volume in 1 Master Mix for four Reagent Stock concentration 10X 25 mM one reaction 1X 2 mM reaction 25 jl reactions Buffer MgClz dNTPs Primer mix DNA template Taq polymerase water Total volume 2.SA 2 al 2.S A Pe 100 nM-0.IA 2 ng/HI 1 unit VS 10 μΜ 0.25 l...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
solve for gel preparation and PCR mastermix calculations with
steps for understanding
TBE Buffer Calculations Determine the mass of the following reagents for a 10X stock 700mM of Tris Base (157g/mol) 887mM of Boric Acid (62g/mol) 25.7mM of EDTA (292g/mol) Dissolve in 750ml of DIH.O and bring to volume (IL) Calculate the dilution of your 10x stock for a 1X working stock. Remember you only need IL of working stock for a single experiment. Gel Preparation Calculations You need to...
2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations for calculating your cocktail. Fill in the chart below. A. Fill in the chart below with the volumes that would be added to each tube. Note the final volume of the reaction. Show your calculations for full credit. REAGENT Final Concentration (50 ul Reaction) 1x Test Reaction (+) Control Reaction 10x Reaction Buffer ML dNTPs (15 mm) 200 UM 0.2 UM ul 0.2 um...
1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteinase K; guanidine HCI; SDS 2. Why is the ratio of the OD at 260 and 280 nm used to estimate DNA purity? 3. In one paragraph, summarize basic principles of PCR technique in your own words. List all the reagents you will need to perform a PCR experiment. Does this method tell you what genetic modifications were made? If yes, describe how you can...
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