YeastLab Report Summary
YeastLab Report Summary
Thereport presents quick and straightforward methods of isolatinggenomic DNA, plasmid DNA, or RNA from yeast. S. cerevisiae isparticularly different from Escherichia coli as its east gene isthree times the size of the genome of the bacterium (Nakamura &Fujimoto, 2013). The genome contains 12 million base pairs which aresufficient to code for about 6000 proteins. Thelab experiment also takes into account all the cell cycle checkpointsand their importance. The functions of different regulators vital inthe cell cycle process. The experiment looks into how the DNAmolecules can be separated by gel electrophoresis (Dalle Nogare,2008). The experiment further highlights the cell growthcharacteristics of S. cerevisiae and identifies the mutant strainusing common biological techniques. Theshape of Saccharomyces cerevisiae is ovoid and contains all theorganelles in a eukaryotic organism (Fong, Arumugam, and Koepp,2013).
Thedifference between S. cerevisiae to other eukaryotic organisms isthat rather than reproducing through binary fission it reproducesthrough budding. The budding process occurs when a small sectionappears on the cell wall or surface and through mitosis one of thenuclei is moved into the bud and it gradually enlarges in size andseparates from the mother cell. Using yeast for the cell cycleexperiment is more prudent because it contains all the standardeukaryotic organelles. It is also a perfect model because of itsbudding process of reproduction. It is also a good choice forexperimental because it has a relatively small genome that has beencompletely sequenced (White & Dalton, 2005).
Thecheckpoints are important since it`s the starting part of the buddingprocess. Each section and part are equally required for the processto be successful. In regards to cancer, the checkpoints are usuallytampered with. Cancer is a result of uncontrolled cell division.Changesin cell cycle regulation are the leading cause of cancer.TheDNA rectification and replacement of damaged DNA happen in the Mphase called G2 checkpoint (Kalli Koniaras et al., 2001).Clb3 andChk1 are regulators in the cell cycle processes. Duringthe experimental period, the Saccharomyces genome deletion occurs,and Chk1 has proteins encoded in the genes (Enserink & Kolodner,2010). Theyare used in the determination of the reproduction process throughbudding and the process of RNA electrophoresis (Winslow &Henkart, 1991).
Electrophoresisis a powerful preparative and analytical technique. Agarose gelelectrophoresis is a safe way to separate RNA and DNA molecules withdifferent sizes. The process begins with a gel of agarose (a purecomponent of agar) is used. It is cast in a flat UV-transparentplastic tray with a Teflon comb positioned near one end.Theextraction of RNA is the purification of RNA molecules in abiological experiment. Thismethod depends on phase separation through centrifugation of amixture of the aqueous sample as well as a solution that containswater-saturated with chloroform and phenol creating an upper aqueousphase and a lower organic phase (Williams, 2002).
ComplementaryDNA can be converted from m RNA. cDNA is a more convenient way towork with the coding sequence than mRNA because RNA is very easilydegraded by omnipresent RNases. PolymeraseChain Reaction (PCR) is used for amplification of specific segmentsof a DNA sample for medical and research purposes (Taylor &Mrkusich, 2014). It is used to obtain enough DNA for genetic testingto test for inherited or acquired mutations like BRCA1/2 or p53 forcancer, CFTR for cystic fibrosis, or CAG trinucleotide repeatexpansion in Huntington’s disease(Hierro et al., 2004).
DalleNogare, D. E. (2008). Molecular-genetic analysis of cell cyclediversification in early zebrafish embryos
Enserink,J. M., & Kolodner, R. D. (2010). An overview of Cdk1-controlledtargets and processes. Cell Division, 5, 11.
Fong,C. M., Arumugam, A., & Koepp, D. M. (2013). The saccharomycescerevisiae F-box protein Dia2 is a mediator of S-phase checkpointrecovery from DNA damage. Genetics, 193(2), 483-99.
Hierro,N., González, Á., Mas, A., & Guillamón, J. M. (2004). NewPCR-based methods for yeast identification. Journal of AppliedMicrobiology, 97(4), 792-801.
KalliKoniaras, Cuddihy, A. R., Christopoulos, H., Hogg, A., &O`Connell, M. J. (2001). Inhibition of Chk1-dependent G2 DNA damagecheckpoint radiosensitizes p53 mutant human cells. Oncogene,20(51), 7453-63.
Nakamura, S.,& Fujimoto, K. (2013). Creation of DNA array structureequipped with heat resistance by ultrafast photocrosslinking. Journalof Chemical Technology & Biotechnology, 89(7),1086-1090. doi:10.1002/jctb.4205
Taylor,S. C., & Mrkusich, E. M. (2014). The state of RT-quantitativePCR: Firsthand observations of implementation of minimum informationfor the publication of quantitative real-time PCR experiments (MIQE).Journal of Molecular Microbiology and Biotechnology, 24(1),46-52.
White,J., & Dalton, S. (2005). Cell cycle control of embryonic stemcells. Stem Cell Reviews, 1(2), 131-8.
Williams,R. (2002). The yeast lifecycle and DNA array technology. Journalof Industrial Microbiology & Biotechnology, 28(3), 186-91.
Winslow,S. G., & Henkart, P. A. (1991). Polyinosinic acid as a carrier inthe microscale purification of total RNA. Nucleic Acids Research,19(12), 3251-3253.