Artificial Intelligence (AI) Calculator for “Nucleic acid extraction yield calculator”
Accurate nucleic acid extraction yield calculation is critical for molecular biology workflows. This calculator quantifies DNA or RNA yield from experimental data.
Explore formulas, tables, and real-world examples to master nucleic acid extraction yield calculations efficiently.
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Example Numeric Prompts for Nucleic Acid Extraction Yield Calculator
- Input: Sample volume = 100 µL, Concentration = 50 ng/µL
- Input: Initial tissue weight = 25 mg, Extraction efficiency = 80%
- Input: Elution volume = 50 µL, Absorbance at 260 nm = 0.8
- Input: RNA concentration = 40 ng/µL, Final volume = 30 µL
Comprehensive Tables of Common Values for Nucleic Acid Extraction Yield
| Sample Type | Typical Yield (ng DNA/mg tissue) | Typical Yield (ng RNA/mg tissue) | Extraction Method | Purity (A260/A280 Ratio) |
|---|---|---|---|---|
| Human Blood (Whole) | 5,000 – 10,000 ng/mL | N/A | Silica Column | 1.8 – 2.0 |
| Mouse Liver Tissue | 10,000 – 20,000 ng/mg | 15,000 – 25,000 ng/mg | Phenol-Chloroform | 1.7 – 1.9 |
| Plant Leaf Tissue | 2,000 – 8,000 ng/mg | 5,000 – 15,000 ng/mg | CTAB Method | 1.8 – 2.1 |
| Bacterial Culture (E. coli) | 20,000 – 50,000 ng/mL | N/A | Alkaline Lysis | 1.8 – 2.0 |
| FFPE Tissue | 500 – 2,000 ng/mg | N/A | Commercial Kits | 1.6 – 1.8 |
| Parameter | Typical Range | Units | Notes |
|---|---|---|---|
| Sample Volume | 10 – 1000 | µL or mL | Depends on sample type |
| Elution Volume | 20 – 100 | µL | Final volume after extraction |
| Concentration | 1 – 1000 | ng/µL | Measured by spectrophotometry or fluorometry |
| Absorbance at 260 nm (A260) | 0.01 – 2.0 | Absorbance Units | Used to estimate nucleic acid concentration |
| Purity Ratio (A260/A280) | 1.6 – 2.1 | Ratio | Indicates protein contamination |
Essential Formulas for Nucleic Acid Extraction Yield Calculation
Understanding the formulas behind nucleic acid extraction yield is fundamental for accurate quantification and downstream applications.
1. Basic Yield Calculation
The total nucleic acid yield (Y) is calculated by multiplying the concentration (C) by the elution volume (V):
- Y: Total nucleic acid yield (ng)
- C: Concentration of nucleic acid (ng/µL)
- V: Elution volume (µL)
Example: If concentration is 50 ng/µL and elution volume is 100 µL, then yield is 5,000 ng.
2. Concentration from Absorbance at 260 nm
Using spectrophotometry, nucleic acid concentration can be estimated from absorbance (A260) using the following formula:
- C: Concentration (ng/µL)
- A260: Absorbance at 260 nm
- D: Dilution factor (unitless)
- ε: Extinction coefficient (ng/µL per absorbance unit)
Common extinction coefficients:
- DNA: 50 ng/µL per A260 unit
- RNA: 40 ng/µL per A260 unit
Example: For DNA, if A260 = 0.8 and dilution factor = 10, then concentration = 0.8 × 10 × 50 = 400 ng/µL.
3. Yield Normalized to Sample Input
To compare yields across samples, normalize total yield to the initial sample input (mass or volume):
- Ynorm: Normalized yield (ng/mg or ng/mL)
- Y: Total nucleic acid yield (ng)
- S: Sample input (mg tissue or mL fluid)
This allows assessment of extraction efficiency relative to starting material.
4. Extraction Efficiency Calculation
Extraction efficiency (E) quantifies the percentage of nucleic acid recovered relative to theoretical maximum:
- E: Extraction efficiency (%)
- Y: Actual yield (ng)
- Ymax: Theoretical maximum yield (ng)
Theoretical maximum yield is often estimated from literature or prior quantifications of nucleic acid content per sample unit.
5. Purity Assessment Using A260/A280 Ratio
Purity is assessed by the ratio of absorbance at 260 nm and 280 nm:
- Ideal DNA purity: 1.8 – 2.0
- Ideal RNA purity: 2.0 – 2.1
- Ratios below 1.6 indicate protein contamination
Real-World Application Cases of Nucleic Acid Extraction Yield Calculation
Case Study 1: DNA Yield from Human Blood Sample
A researcher extracts DNA from 200 µL of human whole blood using a silica column kit. The elution volume is 100 µL. The absorbance at 260 nm is measured as 0.9, and the sample was diluted 1:5 before measurement.
- Calculate the DNA concentration in the eluted sample.
- Calculate the total DNA yield.
- Normalize the yield per mL of blood.
Step 1: Calculate DNA concentration
Using the formula:
Step 2: Calculate total DNA yield
Step 3: Normalize yield per mL of blood
Sample volume = 200 µL = 0.2 mL
This yield is consistent with typical human blood DNA extraction values.
Case Study 2: RNA Extraction from Mouse Liver Tissue
A molecular biologist extracts RNA from 30 mg of mouse liver tissue using phenol-chloroform extraction. The RNA is eluted in 50 µL. The absorbance at 260 nm is 1.2, measured without dilution.
- Calculate RNA concentration.
- Calculate total RNA yield.
- Calculate normalized yield per mg tissue.
- Assess purity if A280 is 0.6.
Step 1: Calculate RNA concentration
Step 2: Calculate total RNA yield
Step 3: Normalize yield per mg tissue
Step 4: Assess purity
A purity ratio of 2.0 indicates high-quality RNA with minimal protein contamination.
Additional Technical Considerations for Nucleic Acid Extraction Yield Calculations
- Instrument Calibration: Spectrophotometers and fluorometers must be calibrated regularly to ensure accurate concentration measurements.
- Sample Dilution: Proper dilution is critical to keep absorbance readings within the linear range (0.1 – 1.0 A260 units).
- Contaminants: Presence of phenol, proteins, or salts can skew absorbance readings; purity ratios help detect contamination.
- Extraction Method Impact: Different extraction protocols yield varying purity and quantity; method selection depends on sample type and downstream application.
- Storage Conditions: Nucleic acid degradation over time affects yield; store samples at -20°C or -80°C for long-term preservation.
- Yield Normalization: Normalizing yield to input sample mass or volume allows comparison across experiments and sample types.
Authoritative Resources and Standards
- NCBI: Nucleic Acid Quantification and Quality Assessment
- Thermo Fisher Scientific: DNA/RNA Purification Handbook
- ISO 20395: Molecular In Vitro Diagnostic Examinations — Nucleic Acid Extraction
- Bio-Rad: Quantification of Nucleic Acids
Mastering nucleic acid extraction yield calculations ensures reliable data for genomics, transcriptomics, and diagnostics.