1. What is the Bioline DNA Concentration Formula?

The Bioline DNA concentration formula estimates double-stranded DNA concentration from absorbance at 260 nm (A260) and the dilution factor. It's based on the principle that 1 A260 unit of dsDNA equals approximately 50 ng/μL of double-stranded DNA.

2. How Does the Calculator Work?

The calculator uses the Bioline DNA concentration formula:

Where:

• \( C \) — DNA concentration (ng/μL)
• \( A_{260} \) — Absorbance at 260 nm (unitless)
• \( DF \) — Dilution factor (unitless)
• 50 — Conversion factor (ng/μL per A260 unit)

Explanation: The formula accounts for the direct relationship between UV absorbance and DNA concentration, with correction for any sample dilution.

3. Importance of DNA Quantification

Details: Accurate DNA quantification is essential for molecular biology applications including PCR, sequencing, and cloning to ensure proper DNA amounts are used in reactions.

4. Using the Calculator

Tips: Enter the A260 reading from your spectrophotometer and the dilution factor used (if any). For undiluted samples, use DF=1.

5. Frequently Asked Questions (FAQ)

Q1: Why is 260 nm used for DNA quantification?
A: DNA absorbs UV light most strongly at 260 nm due to its aromatic nucleotide bases, making this wavelength ideal for quantification.

Q2: What is a good A260 reading?
A: Ideal A260 values are between 0.1-1.0 for accurate measurements. Values outside this range may need dilution or concentration.

Q3: How does purity affect the calculation?
A: The calculation assumes pure DNA. Contaminants like protein or RNA will affect accuracy. Check A260/A280 ratio (should be ~1.8 for pure DNA).

Q4: Does this work for single-stranded DNA or RNA?
A: No, this formula is for double-stranded DNA. For ssDNA use 33 ng/μL per A260 unit, for RNA use 40 ng/μL per A260 unit.

Q5: What if my sample was diluted?
A: Multiply the result by your dilution factor. For example, if you diluted 1:10, enter DF=10.