Fundamental Molecular Biology notes

a. Explain what is a transcriptome b. Differentiate between transient and stable transfection/transformation c. Differentiate between constitutive expression, spatial expression and temporal expression d. Explain what a microarray is e. Explain how a microarray can be used to detect differences in gene expression

a. Explain why due to totipotency it is much easier to produce transgenic plants than transgenic animals b. Discuss the process of tissue culture c. List and discuss the major strategies for gene transfer to plant cells i. Explain agrobacterium transformation in detail by referring to the Ti plasmid, T- DNA, border sequences, vir genes, oncogenes ii. Discuss the experimental protocol for transferring genes via agrobacterium in dicots 2. Discuss electroporation and microballistics (gene gun) as...

a. Compare and contrast three distinct methods for genetic manipulation of organisms b. Discuss step by step how to make a transgenic mouse c. Explain how transgene expression can be analysed d. What is meant by “inducible transgenic mice”? e. What is “knockout mouse” f. Discuss the role of model organisms in Molecular biology studies i. Consider the characteristics of a model organism as well.

a. Describe what DNA sequencing is b. Explain the Sanger sequencing method i. In your answer refer to the primer, type of nucleotides and polymerase c. Read a DNA sequence from an autoradiograph d. Explain how DNA sequencing can be automated by replacing radioactive labels with fluorescent labels e. Explain the use of next-generation sequencing f. Explain the principle of “sequence-by-synthesis”

a. Define electrophoresis b. Explain which molecules can be analysed using electrophoresis c. Explain how an electrical current can separate DNA fragments based on their size d. Explain why smaller fragments move faster through the gel then bigger fragments e. Determine the size of unknown DNA fragments by using a DNA ladder f. Construct a restriction map based on sizes of fragments g. Explain the role of positive and negative controls in an experiment h. Explain the function of a loading d...

a. Explain why plasmids are necessary to produce recombinant DNA b. Explain the difference between chromosomal DNA and plasmid DNA within E.Coli. c. Describe the three different conformations in which plasmid DNA is found d. Describe how these different conformations will separate on an agarose gel during electrophoreses e. Explain how plasmid DNA is purified from E.coli cells by using the alkaline lysis approach. In your answer refer to the function of each reagent f. Explain why the above-me...

a. Explain the role of DNA packaging in gene expression regulation b. Explain by means of an example how DNA packaging can regulated gene expression c. Describe the role of transcription factors, enhancers, silencers and promoters in transcriptional regulation d. Explain how DNA-binding proteins can interact at a distance e. Explain why not all gene regulation targets transcription initiation f. Explain how alternative splicing give rise to different mRNA thus assisting with gene regulation ...

Discuss the sources of DNA for cloning 2. Explain the theoretical principles of PCR and the use of this technique in recombinant genetic technology & biotechnology a. Explain what PCR is b. Explain the role of primers, dNTPs, Taq polymerase and buffer in the reaction c. Describe the three steps of PCR d. Draw the first 3 cycles of PCR

Describe the structure of DNA. a. Describe the primary and secondary structure of DNA b. Discuss alternative double-helical structures c. Discuss denaturation, melting temperature and renaturation d. Describe the tertiary structure of DNA by referring to supercoiling 2. Explain the structure of a chromosome a. Explain the processes involved in the packaging of DNA b. Discuss the role of packaging in a cell 3. Explain how nucleosomes affect nuclear processes to control gene expression 4. De...