Make the PCR Cocktail This table lists the ingredients, stock reagent concentrations, and concentrations in the PCR reaction. Prepare a "PCR cocktail" to be added to your samples to achieve these concentrations. Make enough cocktail to run nine samples. [Four student samples + three positive controls + one negative control + one extra.] {Hint: Remember that the concentration in the reaction is not the same as the concentration in the cocktail!] Component Stock Concentration Concentration in the PCR reaction Volume per reaction Volume to make cocktail Sterile water - - µl µl PCR buffer w/ MgCl2 10x 1x µl µl Nucleotide mix 10 mM 0.2 mM µl µl Primer 1 (Forward) 10 µM 1.0 µM µl µl

Make the PCR Cocktail This table lists the ingredients, stock reagent concentrations, and concentrations in the PCR reaction. Prepare a "PCR cocktail" to be added to your samples to achieve these concentrations. Make enough cocktail to run nine samples. [Four student samples + three positive controls + one negative control + one extra.] {Hint: Remember that the concentration in the reaction is not the same as the concentration in the cocktail!] Component Stock Concentration Concentration in the PCR reaction Volume per reaction Volume to make cocktail Sterile water - - µl µl PCR buffer w/ MgCl2 10x 1x µl µl Nucleotide mix 10 mM 0.2 mM µl µl Primer 1 (Forward) 10 µM 1.0 µM µl µl

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 14QP

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Component	Stock Concentration	Concentration in the PCR reaction	Volume per reaction	Volume to make cocktail
Sterile water	-	-	µl	µl
PCR buffer w/ MgCl₂	10x	1x	µl	µl
Nucleotide mix	10 mM	0.2 mM	µl	µl
Primer 1 (Forward)	10 µM	1.0 µM	µl	µl
Primer 2 (Reverse)	10 µM	1.0 µM	µl	µl
Taq DNA polymerase	5 U/µl	1.0 U	µl	µl
DNA template (sample)	-	~1 ng	20 µl	µl
Total	-	-	40 µl	µl

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Make the PCR Cocktail I field out my work.. but I don't know which one is incoeert.. This table lists the ingredients, stock reagent concentrations, and concentrations in the PCR reaction. Prepare a "PCR cocktail" to be added to your samples to achieve these concentrations. Make enough cocktail to run nine samples. [Four student samples + three positive controls + one negative control + one extra.] {Hint: Remember that the concentration in the reaction is not the same as the concentration in the cocktail!] Component Stock Concentration Concentration in the PCR reaction Volume per reaction Volume to make cocktail Sterile water - - 0µl 0µl PCR buffer w/ MgCl2 10x 1x 4µl 36µl Nucleotide mix 10 mM 0.2 mM 0.8µl 7.2µl Primer 1 (Forward) 10 µM 1.0 µM 4µl 36µl Primer 2 (Reverse) 10 µM 1.0 µM 4µl 36µl Taq DNA polymerase 5 U/µl 1.0 U 8µl 72µl DNA template (sample) - ~1 ng 20 µl 180µl Total - - 40 µl 360 µl
Our PCR samples already contain loading dye, but sometimes this isn’t the case. If your samples didn’t already contain dye and you wanted to load your PCR sample onto an agarose gel, you’d need to add loading dye to the proper concentration. There is a 6X loading dye available for use; how many µl of this loading dye will you add to 10 µl of your sample so that it is at a 1X working concentration? Show your work.
A typical polymerase cycle reaction (PCR) program (Figure 3) followed as: HOLD 1 HOLD 3 HOLD 2 (30CYCLES) 95° C 94°C 72C 72° C 04:00 1:00 1:00 10:00 52.0° C 1:00 04° C 00 Figure 3 (i) Why is polymerase cycle reaction (PCR) repeated 30 times (Figure 3)? What are the differences between primers for PCR program, and primers for DNA replication process during S-phase? (ii) (iii) What are the FIVE (5) basic reagents used in PCR?
PCR primers Below is a 300 base pair fragment of DNA. The top strand is written in the 5' to 3' direction. The bottom strand is written 3' to 5'. There are also two primer sequences; both primers are written 5' to 3'. Note that we are displaying a double-stranded DNA fragment, but primers will only bind to one of the two displayed strands. 5' ACCGȚAGCTATATGCTATCGTGACGTATCGGCGCATTAAȚCGGGATCGAT 3 50 3' TGGCÁTCGATATACOATAGCACTOCATAGCCGCGTAATTÀGCCCTAGCTÀ 5' 5' AGCTÇGCTAGCAGGAGAGAȚATCGÇTCATAGCTCCGATCGATGCCGCTAA 3 3' TCGAGCG ATCGTCCTCTCTÁTAGCGAGTATCGAGÓCTAGCTACGGCGATİ 5' 100 5' TATAGCTCTÇTGCGGATATÇGCATATACCẠ AGGCCCTACGTATGTAGCTA 3 150 3' ATATČGAGAGACOCCTATAGCGTATATGGTTCCGGGATGČATACATCGAŤ 5' 5 TGCGTATATÇGGAGAGTCCTGGATATGGAGCTTGACTGCAGAGAGCTCGA 3 200 3' ACGCÁTATAGCCTCICAGGÁCCTATACCTCGAACTGACGTCTCTCGAGCT 5' 5' TATGCGCTTAGGCCGTATATGCTTGGGGAAAGCTCTATGTATGCTATGTG 3 3. ATACGCGAATCCGGCATATACGAACCCCTÍTCGAGATACATACGATACAC 5' 250 5' TGCATGTGCTATGCAACGTTCOGATTGCGȚAGCAGTAATAGCGCCGATTG 3 300 3'…
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?
The process of PCR essentially revolves around three phases. Briefly describe these phases and the events that occur in them. Take note the temperature on which these phases take place.
What would be the expected effect on the PCR reaction, if you increased the temperature of the annealing phase and the length of the elongation phase? O Accuracy will be reduced, but yield will be increased Accuracy and yield will be increased OAccuracy will be increased, but yield will be decreased Accuracy and yield will be reduced
DNA extraction, PCR, gel electrophoresis, and DNA sequencing of PCR products. Give a description of these methods and the materials required to perform each of these molecular techniques
PCR primers Below is a 300 base pair fragment of DNA. The top strand is written in the 5' to 3' direction. The bottom strand is written 3' to 5'. There are also two primer sequences; both primers are written 5' to 3'. Note that we are displaying a double-stranded DNA fragment, but primers will only bind to one of the two displayed strands. 5' ACCOȚAGCTATATOCTATCOTGACOTATCOGCOCATTAAȚCGGGATCGAT 3 3' TGGCATCGATATACGATAGCACTGCATAGCCGCGTAATTAGCCCTAGCTẢ 5 50 5' AGCTCGCTAGCAGGAGAGATATCGCTCATAGCTCCGATCGATGCCGCTAA 3 100 3' TCGAGCGATCGTCCICTCTATAGCGAGTAICGAGGCTAGCTACGGCGATİ 5' 5' TATAGCTCTCTGCGGATATÇGCATẠTACCAAGGCCCTACGTATGTAGCTA 3 150 3' ATATČGAGAGACGCCTATAGCGTATATGGÍTCCGGGATGČATACATCGAŤ 5 5' TGCGȚATATÇGGAGAGTCCTGGATAT GGAGCTTGACTGCAGAGAGCTCGA 3 200 3' ACGCATATAGCCTCICAGGACCTATACCTCGAACÍGACGICTCTCGAGCİ 5' 5' TATGCGCTTAGGCCGTATATGCTTGGGGAAAGCTCTATGTATGCTATGTG 3 250 3' ATACGCGAATCCGGCATATACGAACCCCTITCGAĞATACATACG ẢTACAČ 5' 5' TGCATOTGCTATOCAACGTTC GGATTGCGȚAGCAGTAATAGCGCCGATTO 3' 300 3'…
In a typical PCR reaction what is happening in stages occurring at temperature ranges (a) 92–95°C, (b) 45–65°C, and (c) 65–75°C.
The final amount of each primer required in a PCR reaction is 25 picomol. If the total volume of the PCR reaction is equal to 100 µl and the stock concentration of each primer is equal to 0.0025 mM. Calculate the volume of stock primer that needs to be added in order to ensure a primer amount of 25 picomol.
This is what the LAB FLOW would look like: CULTURE BACTERIA EXTRACT RNA RANDOM HEXAMER REVERSE TRANSCRIPTION OF RNA PCR OF lacZ GENE ELECTROPHORESIS If you wanted to prepare only one PCR reaction, how much of each reagent would you add to the PCR tube? What would be the final primer concentration if 0.5 μl of 10 μM primers were added to a PCR reaction with a final volume of 20 μl?