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2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations...
Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations for...
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 µl Reaction) Test Reaction (+) Control Reaction (-) 10x Reaction Buffer 1X mL mL dNTPs (15 mM) 200 µM mL mL Forward Primer...
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,...
biochemistry question Nucleic Acids 33 boslari, no NO E Let's Make a PCR Reaction Reaction Mixture:...
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,...
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...
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...
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...
Prepare a fragment of DNA to be cloned by PCR by preparing the oligonucleotides received through...
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...
qRT-PCR The reaction mixture for qRT-PCR requires the following: Taq Mix : 1 uL, SYBR Green...
qRT-PCR The reaction mixture for qRT-PCR requires the following: Taq Mix : 1 uL, SYBR Green Mix: 25 uL, Forward primer : 1 uL, Reverse primer : 1 uL, ROX Reference Dye : 1 uL, Template : 50 ng RNA , Volume up to 50 uL with nuclease free water. If you are going to make a 20 uL reaction mixture and have a 10 ng/uL RNA sample, how much of each of the components do you need? You also...
You are using PCR to amplify a 300 bp target sequence, a portion of Gene X,...
You are using PCR to amplify a 300 bp target sequence, a portion of Gene X, from human genomic DNA isolated from patients' blood samples. The instructions for this procedure tell you to include Samples A and B, whose contents are listed below, with each batch of patient samples that you run. Ingredients Sample A Sample B 10x PCR Buffer (Tris,KCI,MgCl2,BSA) 5 mL 5 mL H2O 37.8mL 38.8mL dNTP's 3 mL 3 mL Taq DNA polymerase 0.2 mL 0.2 mL...
Prepare a PCR reaction using the following recipe. 5x buffer..............................5 μl 25 μM forward primer..........2 μl,...
Prepare a PCR reaction using the following recipe. 5x buffer..............................5 μl 25 μM forward primer..........2 μl, 25 μM reverse primer..........2 μl 5 ng/μl Template..................1 μl 2.5 mM (each) dNTPs..........4 μl Water....................................30 μl Phusion polymerase.............1 μl Q6. What are the final concentrations of the buffer, the forward primer, the template, and the dNTPs in the PCR reaction? Leave all concentrations in the same type of units they were provided in (x, molarity, or weight per volume). Remember the equation C1 x...
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,...
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,...
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...
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...
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...
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...
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