Answer. 6 as given that in one reaction 0.50 ul of template DNA is used. So for 5 reaction 0.50 × 5 = 2.5 ul of template DNA used. Also Total mix is 25 ul then for 5 reaction it will be 25 × 5 = 125 ul.
Also master mix (without template for 1 reaction ) is 24.5 up and master mix for 5 reaction = 122.5 ul.
So, To make individual reactions you would add 24.5 ul master mix to 0.50 ul of the template DNA.
Also for making 5 reaxtion you would add 122.5 ul master mix to 2.5 ul of the template DNA.
How to solve for question#6? 5 5. Fill in the table below to set up the...
4 total reactions 10. Complete this Master Mix table for 3 DNA samples, a positive control, negative control, and an extra reaction for pipetting error. Show your work. 4 pts) Master C Mix Conc. Of Final Stock solution L/Rxn Number of Total L (total per Reactions needed tube) for master mix PCR buffer Water dNTP mix MgCl2 F Primer 20X 1X | | 25 mM2.5 mM 10uM | 0.1 μΜ 100 ml 100 μΜ RPrimer | 10uM | 0.1μΜ Taq...
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,...
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...
Question 5 Using stock solutions in a protocol: from volume to final concentration A reverse transcription reaction is carried out in a 30 μL reaction mix. This type of reaction is carried out to convert RNA to complementary DNA. You are going to set up a number of these reactions so you decide to make up a larger volume of the reaction mixture, called a master mix. This saves on multiple repetitive pipetting and reduces errors. You have been given...
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...
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...
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,...
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...
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...