Home › Biochemistry › Concentration Determination of Nucleic Acids and Proteins Using the Micro-volume Bio-spec Nano Spectrophotometer
Biochemistry JoVE (Open Access) Citable · DOI

Concentration Determination of Nucleic Acids and Proteins Using the Micro-volume Bio-spec Nano Spectrophotometer

DOI: 10.3791/2699-v
What you'll learn
  • ✓ Quantify dsDNA concentration using micro-volume UV-VIS spectrophotometry
  • ✓ Measure protein concentration with minimal sample volume (1-2 µL)
  • ✓ Operate automated wiping and sample handling for reproducible results
  • ✓ Interpret and validate quantitation data for accuracy and carryover
Protocol

Biopharma Insights This communication presents data on the accuracy and reproducibility of the BioSpec-nano UV-VIS spectrophotometer for dsDNA and protein quantitation. Even with ultra-small volumes (1 to 2 L), reproducibility is excellent, while the automated wiping function improves throughput and results in minimal carryover for more precise results.

Difficulty
beginner
Total time
~5-10 min per sample set

Steps

1
Quantify dsDNA using spectrophotometer

Load 1-2 µL dsDNA sample into the BioSpec-nano instrument and measure absorbance at 260 nm to determine nucleic acid concentration. The automated system provides rapid quantitation with minimal sample loss.

▶ 00:54
2
Quantify protein using spectrophotometer

Load 1-2 µL protein sample and measure absorbance at 280 nm to determine protein concentration. Use the instrument's protein-specific algorithms for accurate quantitation of small volumes.

▶ 03:27
3
Validate quantitation results and carryover

Review representative output data for DNA and protein samples, confirming reproducibility across replicates and verifying minimal carryover between consecutive measurements via the automated wiping function.

▶ 04:37
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