1. What is the Bioline Calculation Tool?

The Bioline Calculation Tool estimates DNA/RNA concentration from absorbance measurements using the Beer-Lambert law. It provides a quick and accurate way to determine sample concentration based on spectrophotometer readings.

2. How Does the Calculator Work?

The calculator uses the basic formula:

Where:

• \( Absorbance \) — Measured absorbance at 260nm (AU)
• \( Extinction\ Coefficient \) — Substance-specific constant (ml/μg cm)

Explanation: The equation relates the amount of light absorbed by a solution to the concentration of the absorbing species in that solution.

3. Importance of Concentration Calculation

Details: Accurate concentration measurement is crucial for molecular biology experiments, ensuring proper DNA/RNA quantities for downstream applications like PCR, sequencing, and cloning.

4. Using the Calculator

Tips: Enter absorbance reading (typically at 260nm) and the appropriate extinction coefficient for your nucleic acid. For dsDNA, the standard coefficient is 0.05 ml/μg cm.

5. Frequently Asked Questions (FAQ)

Q1: What is a typical extinction coefficient for DNA?
A: For double-stranded DNA: 0.05 ml/μg cm. For single-stranded DNA or RNA: 0.033 ml/μg cm.

Q2: What absorbance wavelength should I use?
A: For nucleic acids, use 260nm. The 260/280 ratio (using 280nm) helps assess purity.

Q3: How do I measure absorbance?
A: Using a spectrophotometer with the sample in a quartz cuvette (plastic may absorb UV light).

Q4: What affects absorbance readings?
A: Sample turbidity, buffer composition, and contaminants can affect accuracy.

Q5: Is this calculator suitable for proteins?
A: No, proteins use different wavelengths (280nm) and extinction coefficients.