Isolation of Yeast Genomic DNA

Isolating genomic DNA from yeast involves culturing the microbe, harvesting the cell, enzymatically removing the cell wall, lysing the protoplast, and finally separating the DNA from the other cell debris.
Materials that you need are:

Yeast culture-prepared previously

Spectrophotometer with cuvettes

50 mM EDTA, pH 8-ice cold

50 mM Tris, pH 9.5, 2% 2-mercaptoethanol

1.2 M sorbitol, 50 mM Tris, pH 7.5

Lyticase solution-500 U/ml in 50 mM Tris, pH 7.5

10% Sodium Dodecyl Sulfate (SDS)-used for checking protoplast formation

Lysis buffer-100 mM Tris, pH 7.5, 100 mM EDTA, 150 mMNaCl, 50 micrograms/ml RNase A

Lysis buffer with 2% SDS

95% Ethanol-stored at minus 20 degree Celcius

TE buffer-10 M Tris, pH 8, 1 mM EDTA

3 M potassium acetate, pH 5.5

Here are the step by step methods:

Bacterial DNA Extraction using InstaGene® Matrix

In spite of using Phenol and Chloroform to extract bacterial DNA template, you can use the InstaGene matrix to be more efficient. This DNA extraction method is very easy, fast, and practical. Here is the procedure:

References

Here are the references of the blog:

1. Prieto, L.M. et al., “The production of rhamnolipid by a Pseudomonas aeruginosa strain isolated from a southern coastal zone in Brazil”, Chemosphere (2008), doi:10.1016/j.chemosphere.2008.01.003.
2. Burden, D.W.; Whitney, D.B., "Biotechnology, Proteins to PCR, A Course in Strategies and Lab techniques", Birkhauser Boston, 1995.
3. Thaniyavarn, J. et al., "Production and Characterization of Biosurfactants from Bacillus licheniformis F2.2" , Biosci. Biotechnol. Biochem., 67 (6), 1239-1244, 2003
4. Eeles, R.A.; Stamps, A.C., "Polymerase Chain Reaction (PCR) The Technique And Its Applications", The Institute of Cancer Research, The Royal Marsden Hospital, Sutton, Surrey, United Kingdom, 1993.
5. Aitken, A.; Learmonth, M., "The Protein Protocols Handbook, Second Edition: Protein Determination by UV Absorption", Humana Press, 2002.
6. Aitken, A.; Learmonth, M., "The Protein Protocols Handbook, Second Edition: Quantitation of Tryptophan in Proteins ", Humana Press, 2002.
7. Heptinstall. J and Rapley. R, “The Nucleic Acid Protocols Handbook: Spectrophotometric Analysis of Nucleic Acids”, Humana Press, 2000.
8. Walker, J.M., "The Protein Protocols Handbook, Second Edition: The Bicinchoninic Acid (BCA) Assay for Protein Quantitation", Humana Press, 2002.
9. Waterborg, J.H., "The Protein Protocols Handbook, Second Edition: The Lowry Method for Protein Quantitation", Humana Press, 2002.
10. Kruger, N.J., "The Protein Protocols Handbook, Second Edition: The Bradford Method for Protein Quantitation", Humana Press, 2002.
11. Kolmodin, L. A, Williams, J.F, “The Nucleic Acid Protocols Handbook: Polymerase Chain Reaction, Basic Principles and Routine Practice”, Humana Press Inc., Totowa, NJ, 2000.
12. Chachaty. E, Saulnier. P, “The Nucleic Acid Protocol Handbook: Bacterial DNA Extraction for Polymerase Chain Reaction and Pulsed-Field Gel Electrophoresis”, Humana Press Inc., Totowa, NJ, 2000.
13. Williams. D. R, Rapley. R, “The Nucleic Acid Protocol Handbook: Agarose Gel Electrophoresis of Nucleic Acids”, Humana Press Inc., Totowa, NJ, 2000.
14. Smith. D. R, “The Nucleic Acid Protocol Handbook: Restriction Endonuclease Digestion of DNA”, Humana Press Inc., Totowa, NJ, 2000.
15. Stewart, T.L., Mann, V., “Methods in Molecular Medicine, Vol. 80: Bone Research Protocols”, Humana Press Inc., Totowa, NJ, 2003.
16. Theophilus, B. D. M., “Methods in Molecular Biology, Vol. 86: RNA Isolation and Characterization Protocols”, Humana Press Inc., Totowa, NJ, 1998.
17. Perry, J., Parniske, M., "Lotus japonicus Handbook: 96-WELL DNA ISOLATION METHOD", 2005 Springer.
18. Winstanley, C., Rapley, R., “The Nucleic Acid Protocols Handbook: Extraction and Purification of Plasmid DNA”, Humana Press Inc., Totowa, NJ, 2000.
19. Bogner, P., Killeen, A. A., “Molecular Diagnostics: For the Clinical Laboratorian, Second Edition: Extraction of Nucleic Acids”, Humana Press Inc., Totowa, NJ, 2005.
20. Harwood. A. J, “The Nucleic Acid Protocol Handbook: Native Polyacrylamide Gel Electrophoresis”, Humana Press Inc., Totowa, NJ, 2000.
21. Aitken. A, Learmonth. M, “The Protein Protocols Handbook, Second Edition: Estimation of Disulfide Bonds Using EIIman's Reagent”, Humana Press Inc., Totowa, NJ, 2002.
22. Miller. R. M, Zhang. Y, “Methods in Biotechnology, Vol 2 Bioremediation Protocols: Measurement of Biosurfactant-Enhanced Solubilization and Biodegradation of Hydrocarbons”, Humana Press Inc., Totowa, NJ, 1997.
23. Boerner. S. A, Lee. Y. K, Kaufmann. S. H, Bible. K. C., “The Protein Protocols Handbook, Second Edition: The Nitric Acid Method for Protein Estimation in Biological Samples", Humana Press, 2002.

How to measure emulsification index (E24)

In biotechnology field, this method is used to measure the emulsification ability of biosurfactant. Emulsification index (E24) is used to characterize the biosurfactant in emulsifying, generally, the hydrophobic phase in hydrophilic phase.

Following s are the emulsification index procedure based on the published journal titled “The production of rhamnolipid by a Pseudomonas aeruginosa strain isolated from a southern coastal zone in Brazil” by L.M. Prieto, et al.:

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