Safety in Chemical Laboratories. Introduction 1. A chemical lab is potentially hazardous environment...
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Transcript of Safety in Chemical Laboratories. Introduction 1. A chemical lab is potentially hazardous environment...
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Safety in Chemical Safety in Chemical LaboratoriesLaboratories
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IntroductionIntroduction1. A chemical lab is potentially
hazardous environment2. Accident and injury can happen
anytime3. Lab safety is everyone’s
responsibility4. Lab safety standards and
practices must be strictly followed
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Fire ExtinguisherFire Extinguisher
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DNA Extraction and DNA Extraction and PurificationPurification
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Lab EquipmentsLab Equipments(To be used in this experiment)(To be used in this experiment)
Automatic pipettes
Microcentrifuge
Vortex
Water bath
UV-spectrophotometer
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Steps in DNA Extraction Steps in DNA Extraction and Purificationand Purification
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Quantification of the Quantification of the purified DNApurified DNA
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SpectrophotometerSpectrophotometerMost of visible spectrophotometers are
composed of:◦ Light source which works with visible
wavelengths (400-700 nm)
◦ Monochromator filter for choosing desired wavelength
◦ Sample holder (cuvette)◦ Detector◦ Meter or recorder
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MeasurementsMeasurements◦Dilute the sample of purified DNA:
Add 700 μL of AE to the purified DNA ◦Measure the Absorbance at 260nm◦ Measure the Absorbance at 280nm◦ Assess the DNA purity: 260/280 ratio
(Accepted ratio: 1.7 - 1.9)◦ Calculate DNA Conc.: Provided A260 = 1.0,
DNA is 50 μg/ml, unknown DNA conc. can be calculated by cross multiplicationA260 = 1.0 DNA conc. = 50 μg/mlA260 = 0.5 DNA conc. ? Calculate DNA Yield: DNA Volume x DNA conc.
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DNA Yield:DNA Yield:
◦ If you have Volume of DNA solution: 100μl (0.1 ml) DNA conc.: 30 μg/ml
Then the yield (μg) = Volume x Conc. 0.1 ml x 30 μg/ml = 3.0 μg
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DNA ApplicationsDNA Applications Purified DNA can be used for:
1. Molecular diagnosis of diseases (e.g., sickle cell anemia)
2. Forensic applications (e.g., paternity testing)
3. Molecular biology research
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Molecular techniques using purified DNA:
a. Amplification techniques: polymerase chain reaction (PCR)
b. Southern blottingc. Restriction Fragment length polymorphism (RFLP)