This application note describes in detail how the Agilent Technologies 2100 bioanalyzer can be used to analyze PCR fragments using the DNA 7500 LabChip® kit. Both size and concentration information can be obtained for fragments ranging from 100–7500 base pairs. Sizing and quantitation accuracy were determined and performance was compared to slab gel electrophoretic methods. The advantages of the DNA 7500 assay for determining PCR product purity are also described.
This document details the use of the Agilent 2100 Bioanalyzer to perform quantitative analysis of PCR fragments using the DNA 7500 LabChip kit. It compares the performance of this chip-based system against traditional slab gel electrophoresis, highlighting advantages in speed, sensitivity, and data precision. The note covers the analysis of fragments ranging from 100 to 7500 base pairs.
| Fragment size range | 100-7500 base pairs |
|---|---|
| Sizing accuracy | better than 85 % bp |
| Quantitation accuracy | better than 70 % |
| Linear range of quantitation | 0.5 - 50 ng/pl |
| Analysis time | less than 30 minutes |
| Detection wavelength | 670 nm and 700 nm |
The linear range of quantitation is 0.5 to 50 ng/pl.
No, the assay is compatible with commonly used PCR buffers so no desalting or other sample pre-treatment is necessary.
The Agilent 2100 bioanalyzer analyzes 12 DNA samples in a sequential manner.