Question

In: Biology

You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR...

  1. You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR primers that should amplify a 1 kilobase pair (kbp) PCR product that contains the gene of interest.  After amplification, you will see if the PCR  was successful by loading the entire reaction onto an agarose gel and performing electrophoresis to see if a product of the expected size was generated.  To visualize the DNA, you will stain the gel with a fluorescent dye called ethidium bromide, which fluoresces when it binds to DNA. The sensitivity of ethidium-bromide-stained DNA is 10 nanograms (i.e. – there must be at least 10 ng of DNA in the band in the gel to emit a detectable amount of light).

If your PCR reaction initially contained 30 B. sanfranciscus genomes, how many cycles of PCR will required before there is a detectable amount of amplified product?  You can assume:  a) there is 1 copy of the gene per genome, b) the PCR occurs with perfect efficiency and therefore the amount of product doubles after each cycle, and, c) that the molecular weight of a 1 kbp molecule of DNA is 6.5 x 105Daltons. Express your answer in the number of complete (not fractional) cycles and show your work.

Solutions

Expert Solution


Related Solutions

During “cut and paste” cloning, the PCR-amplified gene is cut by restriction enzymes. How does the...
During “cut and paste” cloning, the PCR-amplified gene is cut by restriction enzymes. How does the synthesis of DNA oligonucleotide primers facilitate the ability to ‘cut’ the amplified gene?
You are setting up a PCR reaction using DNA from a bacteria with a genome size...
You are setting up a PCR reaction using DNA from a bacteria with a genome size of 17 megabase-pairs (mb). How many nanograms of DNA will you need to add to the reaction to ensure there are at least 100 copies of the single gene you are trying to amplify? (assume the average molecular weight of a nucleotide pair is 650). (2 pt)
Describe the principles behind and the applications of the following: a) Reverse transcriptase-PCR b) Cloning DNA...
Describe the principles behind and the applications of the following: a) Reverse transcriptase-PCR b) Cloning DNA into a plasmid vector c) SDS-PAGE d) Restriction mapping e) Sanger Sequencing of DNA Note Detail explanation is required for each .
Design an experiment to knock out a certain gene of interest in the human genome. Outline...
Design an experiment to knock out a certain gene of interest in the human genome. Outline the experiment, explaining the methods you would use, and also indicate how you would verify your experiment was successful at each step.
Differentiate genome reduction from horizontal gene transfer. Specifically in bacteria.
Differentiate genome reduction from horizontal gene transfer. Specifically in bacteria.
A student is tasked with cloning a specific gene-coding sequence from a human cancer cell in...
A student is tasked with cloning a specific gene-coding sequence from a human cancer cell in order to produce the corresponding protein in bacteria at a concentration sufficient to conduct in vitro biochemical studies. B) Another student mentions that her should have cloned the gene sequence using mRNA instead of genomic DNA. Is this a good suggestion? How does one go about doing this?
You are interested in determining the heat of isomerization of two isomers (A and B), so...
You are interested in determining the heat of isomerization of two isomers (A and B), so you measure their heats of combustion. The difference between the heats of combustion is equal to the heat of isomerization. You are given two separate samples of (1.5 ± 0.1) mol for each isomer, A and B, and they are burned in a constant-pressure calorimeter. The heat of capacity of the calorimeter is (8.44 ± 0.05) kJ K-1. Combustion of isomer A causes the...
. What is an mRNA? 2. How does the RT-PCR tell you if a gene is...
. What is an mRNA? 2. How does the RT-PCR tell you if a gene is expressed in a particular cell type? 3. Where can RNases be found? How can you prevent RNase contamination in your samples during experiments involving RNA?
During the analysis of the genomic sequence of the mouse genome, you have found a gene...
During the analysis of the genomic sequence of the mouse genome, you have found a gene that you suspect may be involved in resistance of infection to bubonic plague. You want to test your suspicions by creating a knockout mouse. Outline the steps you would have go through to create such a mouse that is homozygouse for disruption of both copies of this gene. Plese include terms like ES cells, neomycin, positive-negative selction, homologous recombination, blastocyst, and chimera.
3) How can you use PCR to clone a 2000bp gene into a plasmid vector?
3) How can you use PCR to clone a 2000bp gene into a plasmid vector?
ADVERTISEMENT
ADVERTISEMENT
ADVERTISEMENT