Share This Article:

The Use of DNA Extraction for Molecular Biology and Biotechnology Training: A Practical and Alternative Approach

Full-Text HTML XML Download Download as PDF (Size:1543KB) PP. 762-772
DOI: 10.4236/ce.2015.68079    4,945 Downloads   6,884 Views   Citations

ABSTRACT

DNA extraction and polymerase chain reaction are molecular biology techniques widely used in research, diagnostics and biotechnology industry. In this work, we report a practical course designed for undergraduate students of health, biological sciences and biotechnology areas. The course is five days long and emphasizes basic procedures in practical activities. The students were challenged to use three different protocols for DNA isolation (one using commercial silica column and two, in-house) using no lab animals but themselves. They evaluated the DNA yield, and potential amplification of the extracted DNA by PCR using their own cells genetic material. Students were stimulated to adopt a critical view during the course to develop skills including critical analysis of primary results and possible errors, scientific report writing and a problem solving profile.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Lázaro-Silva, D. , De Mattos, J. , Castro, H. , Alves, G. and Amorim, L. (2015) The Use of DNA Extraction for Molecular Biology and Biotechnology Training: A Practical and Alternative Approach. Creative Education, 6, 762-772. doi: 10.4236/ce.2015.68079.

References

[1] Aidar, M., & Line, S. R. (2007). A Simple and Cost-Effective Protocol for DNA Isolation from Buccal Epithelial Cells. Brazilian Dental Journal, 18, 148-152.
[2] Azer, S. A., Guerrero, A. P., & Walsh, A. (2013). Enhancing Learning Approaches: Practical Tips for Students and Teachers. Medical Teacher, 35, 433-443.
[3] Badyal, D. K., & Desai, C. (2014). Animal Use in Pharmacology Education and Research: The Changing Scenario. Indian Journal of Pharmacology, 46, 257-265.
[4] Cardoso, C. V. P., & Almeida, A. E. C. C. (2014). Laboratory Animal: Biological Reagent or Living Being? Brazilian Journal of Medical and Biological Research, 47, 19-23.
[5] Coil, D., Wenderoth, M. P., Cunningham, M., & Dirks, C. (2010). Teaching the Process of Science: Faculty Perceptions and an Effective Methodology. CBE-Life Sciences Education, 9, 524-535.
[6] González, C. I., Ortiz, S., & Solari Colombian, A. (2010). Trypanosoma Cruzi Major Genotypes Circulating in Patients: Minicircle Homologies by Cross-Hybridization Analysis. International Journal for Parasitology, 40, 1685-1692.
[7] Gryson N. (2010). Effect of Food Processing on plant DNA Degradation and PCR-Based GMO Analysis: A Review. Analytical and Bioanalytical Chemistry, 396, 2003-2022.
[8] Hearn, R. P., & Arblaster, K. E. (2010). DNA Extraction Techniques for Use in Education. Biochemistry and Molecular Biology Education, 3, 161-166.
[9] Marchese, A., Esposito, S., Coppo, E., Rossi, G. A., Tozzi, A., Romano, M., Da Dalt, L., Schito, G. C., & Principi, N. (2011). Detection of Streptococcus pneumoniae and Identification of Pneumococcal Serotypes by Real-Time Polymerase Chain Reaction Using Blood Samples from Italian Children ≤5 Years of Age with Community-Acquired Pneumonia. Microbial Drug Resistance, 20, 1-6.
[10] Mello, F. C., Souto, F. J., Nabuco, L. C., Villela-Nogueira, C. A., Coelho, H. S., Franz, H. C., Saraiva, J. C., Virgolino, H. A., Motta-Castro, A. R., Melo, M. M., Martins, R. M., & Gomes, S. A. (2007). Hepatitis B Virus Genotypes Circulating in Brazil: Molecular Characterization of Genotype F Isolates. BMC Microbiology, 23, 103.
[11] Miller, S. A., Dykes, D. D., & Polesky, H. F. (1988). A Simple Salting out Procedure for Extracting DNA from Human Nucleated Cells. Nucleic Acids Research, 16, 1215.
http://dx.doi.org/10.1093/nar/16.3.1215
[12] Miller, M. B. (1994). Practical DNA Technology in School. Journal of Biological Education, 28, 203-211.
http://dx.doi.org/10.1080/00219266.1994.9655393
[13] Mortarino, M., Garagiola, I., Lotta, L. A., Siboni, S. M., Semprini, A. E., & Peyvandi, F. (2011). Non-Invasive Tool for Foetal Sex Determination in Early Gestational Age. Haemophilia, 17, 952-956.
http://dx.doi.org/10.1111/j.1365-2516.2011.02537.x
[14] Nomura, M., Shigematsu, H., Li, L., Suzuki, M., Takahashi, T. et al. (2007). Polymorphisms, Mutations, and Amplification of the EGFR Gene in Non-Small Cell Lung Cancers. PLoS Medicine, 4, e125.
http://dx.doi.org/10.1371/journal.pmed.0040125
[15] Poh, J. J., & Gan, S. K. (2014). Comparison of Customized Spin-Column and Salt-Precipitation Finger-Prick Blood DNA Extraction. Bioscience Reports, 34, 629-634.
http://dx.doi.org/10.1042/BSR20140105
[16] Rogers, N. L., Cole, S. A., Lan, H. C., Crossa, A., & Demerath, E. W. (2007). New Saliva DNA Collection Method Compared to Buccal Cell Collection Techniques for Epidemiological Studies. American Journal of Human Biology,19, 319- 326.
http://dx.doi.org/10.1002/ajhb.20586
[17] Salazar, L. A., Hirata, M. H., Cavalli, S. A., Machado, M. O. et al. (1998). Optimized Procedure for DNA Isolation from Fresh and Cryopreserved Clotted Human Blood Useful in Clinical Molecular Testing. Clinical Chemistry, 44, 1748-1450.
[18] Sambrook, J., Fritsch, E. R., & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd ed.). Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
[19] Schijman, A. G., Bisio, M., Orellana, L., Sued, M., Duffy, T., Mejia Jaramillo, A. M. et al. (2011). International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Neglected Tropical Diseases, 5, e931.
http://dx.doi.org/10.1371/journal.pntd.0000931
[20] Slonczewski, J. L., & Marusak, R. (2004). A Response to BIO 2010: Transforming Undergraduate Education for Future Research Biologists, from the Perspective of the Biochemistry and Molecular Biology Major Program at Kenyon College. Biochemistry and Molecular Biology Education, 32, 151-155.
http://dx.doi.org/10.1002/bmb.2004.494032030342
[21] Sitaraman, R. (2012). From Bedside to Blackboard: The Benefits of Teaching Molecular Biology within a Medical Context. Perspectives in Biology and Medicine, 55, 461-466.
http://dx.doi.org/10.1353/pbm.2012.0030
[22] Takayama, T., Yamada, S., Watanabe, Y., Hirata, K., Nagai, A., Nakamura, I., Bunai, Y., & Ohya, I. (2001). Origin of DNA in Human Serum and Usefulness of Serum as a Material for DNA Typing. Legal Medicine, 3, 109-113.
http://dx.doi.org/10.1016/S1344-6223(01)00018-9
[23] Tsui, N. B. Y., Kadir, R. A., Chan, K. C. A., Chi, C., Mellars, G., Tuddenham, E. G., Leung, T. Y., Lau, T. K., Chiu, R. W. K., & Dennis Lo, Y. M. (2011). PCR Analysis of Maternal Plasma DNA Noninvasive Prenatal Diagnosis of Hemophilia by Microfluidics Digital. Blood, 117, 3684-3691.
http://dx.doi.org/10.1182/blood-2010-10-310789
[24] Tibell, L. A., & Rundgren, C. J. (2010). Educational Challenges of Molecular Life Science: Characteristics and Implications for Education and Research. CBE Life Sciences Education, 9, 25-33.
[25] Voet, J. G., Bell, E., Boyer, R., Boyle, J., O’Leary, M., & Zimmerman, J. K. (2003). Recommended Curriculum for a Program in Biochemistry and Molecular Biology. Biochemistry and Molecular Biology Education, 31, 161-162.
http://dx.doi.org/10.1002/bmb.2003.494031030223

  
comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.