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- 7) Why are the following reagents used? Neutralizing solution (Plasmid isolation) Isopropanol (Plasmid isolation) RNase (isolation of genomic DNA)List the antibiotics that could successfully be used to test for the presence of the 4. gene on your plasmid.9. During nucleic acid hybridization, the probe is labelled for DNA stability to increase probe-test DNA binding to identify the location of probe and the test DNA binding for amplification 11. What is not true for Sequence tagged site (STS) markers: cannot be mapped by fluorescence in situ hybridization (FISH) subset of STS markers are known as expressed sequence tag (EST) markers can readily be screened by a PCR assay short DNA sequences that occur at a unique location in the genome
- Discuss two reasons you need to cut the plasmids prior to inserting them into the fertilized egg.7. Consider the following plasmid (size 6700 bp), with restriction sites at the positions indicated: BamHI + 1 6700 bp 2800 3500 EcoRI BamHI Probebiotechnology lab class :1. The bacterial streaking on antibiotic containing agar plates has been completed and the plates will be available for you to inspect in the lab class.2. You will complete a restriction enzyme digest on the four available plasmid stocks and then examine the products by agarose gel electrophoresis. Photographs of the gel are attached to identify the plasmid present in each analysed sample.Now consider the following questions :- For each of the four plasmids used in the practical, identify the size (in kb) of the linear DNA fragment(s) that you would expect to obtain if the EcoRI digest is complete.- Using the provided negative image of the gel, which shows the bands present in the ND and D samples for each plasmid, identify the plasmid that was present in each of the four plasmid stock.- Explain how you were able to identify plasmid in each sample using the gel, considering how linear DNA fragments of different length are separated by agarose gel…
- Explain how and why PCR can be used to amplify DNA. Describe the steps in the process. Be sure to address the role of Taq DNA polymerase.Explain 2 possible consequences of accidentally puncturing one of the wells in the agarose gel in an electrophoresis set-up when loading a DNA sample.1. Compare Illumina/Pacbio/Nanopore sequencing methods.2. How do you construct the profile matrix? Answer all the part in details and with proper examples.
- 1. Briefly describe how you extract and purify the pJET plasmid from the bacteria.1. What is the purpose of restriction enzyme? 2. What is the purpose of the probe in DNA finger printing. 3. What is/are the sample that can be used for DNA finger printing? 4. Who is the culprit base on our interactive laboratory experiment?2. PCR analysis practice One 1) If you collect samples from crime scene and extract DNA out of each sample. Describe the technologies used to get the below results. 2) Identify which suspect is at the crime scene 3) Among suspect 1-3, who is homozygous for the tested locus and who is heterozygous? DNA from crime scene DNA ladder Suspect DNA #1 #2 #3 500 bp 400 bp 300 bp 200 bp 100 bp | |