You've ordered and received your "primers" for your PCR. Now what? In your new role as an MLA your boss asks you to resuspend each DNA oligonucleotide in water to a final concentration of 100 uMolar (100 µM) What volume of water, in ul, do you need to add to the tube on the left? The information about the yield is on the tube label (right) and I've retyped it below to make it clearer. You may, odmay not, need all of it. 231076243XXIDT C.DOLPHIN 450581747 4830 A CAA TOC TAt A A CA 0.1C 4300-27.1ms.2g 23107 231076243 XIDT C.DOLPHIN 450581747 4830 24Jen2 AC CAA T TACA ADA MW7,321.8/mol 63 00- 27.1 nmol-0.20mg

You've ordered and received your "primers" for your PCR. Now what? In your new role as an MLA your boss asks you to resuspend each DNA oligonucleotide in water to a final concentration of 100 uMolar (100 µM) What volume of water, in ul, do you need to add to the tube on the left? The information about the yield is on the tube label (right) and I've retyped it below to make it clearer. You may, odmay not, need all of it. 231076243XXIDT C.DOLPHIN 450581747 4830 A CAA TOC TAt A A CA 0.1C 4300-27.1ms.2g 23107 231076243 XIDT C.DOLPHIN 450581747 4830 24Jen2 AC CAA T TACA ADA MW7,321.8/mol 63 00- 27.1 nmol-0.20mg

Biology Today and Tomorrow without Physiology (MindTap Course List)

5th Edition

ISBN:9781305117396

Author:Cecie Starr, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Christine Evers, Lisa Starr

Chapter10: Biotechnology

Section: Chapter Questions

Problem 10SQ

See similar textbooks

Similar questions

After you did the amplification by PCR, you run the product on 2% gel. Tell me how did you prepared the gel if the tray is 150 ml? how many milliliters of the TBE and how many grams of the Agarose? Draw a picture (On a paper or on computer) for the gel that you are expecting, the gene that you amplified is approximately 200 bp
A student carries out PCR using the following steps: step 1: 94C for 1 minuteStep 2: 60C for 30seconds Step 3: 72C for 30 seconds The student is examining a 1000-bp DNA fragment as part of their research project. If the limit of detection of this molecule 9 x 108 molecules, what is the minimum number of PCR cycles you would have to run to detect PCR product generated from a single molecule of the template? 2. Two double-stranded fragments of DNA are exactly the same length. At 89 C, fragment A has completely denatured which means the two strands have separated. At that temperature, fragment B is still double-stranded. How might these fragments differ to result in different denaturation temperatures?
A student is trying to add 15.0 ng of DNA template to a 20.0 µL PCR. The DNA template is at a concentration of 65.0 ng µL-1, and the student determines that a serial dilution is required because directly adding the DNA template would require a volume less than 1.00 µL. The student wants to prepare an intermediate solution at a concentration of 15.0 ng µL-1. If the DNA template stock will be mixed with 13.0 µL of ultrapure H2O, calculate the volume (in µL) of the DNA template required to prepare the intermediate solution.
You wish to amplify a segment of DNA from a plasmid template by PCR with the use of the following primers: 5’-GGATCGATGCTCGCGA3' and 5' -AGGATCGGGTCGCGAG-3'. Despite repeated attempts, you fail to observe a PCR product of the expected length after electrophoresis on an agarose gel. Instead, you observe a bright smear on the gel with an approximate length of 25 to 30 base pairs. Explain these results.
You want to amplify the following sequence of DNA from a gene of interest that you are studying. To do this, you will use PCR. Design a forward and reverse primer. Show where the primers anneal to the template sequence. Template DNA: 5’ ATGACGGAATATAAGCTGGTGGTGGTGG---GGCTGCATGAGCTGCAAGTGTGTGCTCTCCTAA 3’ 3’ TACTGCCTTATATTCGACCACCACCACC---CCGACGTACTCGACGTTCACACACGAGAGGATT 5’ (PLEASE EXPLAIN STEP - BY STEP) Answer : Foward primer 5": Reverse primer 3"
Look at each PCR component listed below. For each one, determine which steps(s) of the PCR reaction (denaturation, annealing or extension) would be directly affect if that component were missing. Taq polymerase: Oligonucleotide primers: DNA template: Deoxynucleotides (A, T, G and C): Imagine that you correctly prepare your PCR reaction mixture, but there is something wrong with the thermal cycler. Describe what would happen if: The thermal cycler was stuck on 940C: The thermal cycler cycled between 600C and 720C, but never reached 940C The thermal cycler cycled between 940C and 720C, but never reached 600
If you are trying to set up a PCR with a total volume of 40uL and you have access to a stock solution of 5x buffer, how much 5x buffer will you be adding to the PCR? ___uL?
Ben used the following primer pairs to amplify the COI region of his DNA template: Afterwards, he used Agarose Gel Electrophoresis to visualize the PCR products. Based on the gel: a) Was he able to successfully amplify the desired region in all replicates? b) Are there errors in Ben's gel? Does he need to repeat his PCR?
.How much PCR product (500 bp) should be added to a ligation in which 50 ng of Vector (6.6 kb) vector will be used if a 3:1 insert:vector molar ratio is desired? Vector concentration is 50 ng/uL and insert concentration is 35 ng/ul. Provide mass and volumes for the ligation reaction
Your graduate advisor asks you to amplify the following sequence of DNA by PCR: 5’-ATACGCATTCGGACCAGGTCCTAA-3’ 3’-TATGCGTAAGCCTGGTCCAGGATT-5’ a. To ensure that the entire sequence above is amplified, what 6-nucleotide DNA primers should you add to your PCR mix? You order the primers listed above, but instead receive the following set of primers: 5’-CGCATT-3’ 5’-GGACCT-3’ b. What portion of the double stranded DNA molecule will be amplified after 10 rounds of PCR? Your labmate attempts to rescue your PCR reaction by providing you with the following set of primers: 5’-ATACGC-3’ 5’-TCCTAA-3’ c. What is the result of running the PCR reaction with your labmate’s primers? How many double stranded molecules of DNA will result from 10 rounds of amplification?
A student is trying to add 9.0 pmol of primer mix to a 20.0 µL PCR. The primer mix is at a concentration of 50.0 µM, and the student determines that a serial dilution is required because directly adding the primer mix would require a volume less than 1.00 µL. The student takes 1.40 µL of the primer mix stock and mixes it with 25.0 µL of ultrapure H2O. Calculate the volume (in µL) of the intermediate solution required to add the correct amount of primer mix to the PCR.
What primers will be used to sequence your PCR product where are those sequences on your PCR product How did they get there?