GB/T 37873-2019 in English

Analysis of quality evaluation methods

The standard specifies the specific steps from sample preparation to detection, including purity detection, total amount detection, integrity detection and sequence consistency detection. The following is a detailed interpretation of the key detection methods:

1. Purity detection

The spectrophotometric method was used to determine $\mathrm{OD}_{260}$, $\mathrm{OD}_{280}$ and $\mathrm{OD}_{230}$, and the DNA purity was evaluated by ratio analysis. High-purity DNA should satisfy $\mathrm{OD}_{260}/\mathrm{OD}_{280} \in [1.8, 2.0]$ and $\mathrm{OD_{260}/OD_{230}} \geq 2.0$.

2. Total mass detection

Measure $\mathrm{OD}_{260}$ with a spectrophotometer, and calculate the mass concentration and total mass of DNA using the formula. The calculation formula is as follows:

Wherein, $N$ is the sample dilution multiple, $F$ is the double-stranded DNA conversion factor ($50~\mu\mathrm{g}/\mathrm{mL}$), and $V$ is the sample volume.

3. Integrity test

Agarose gel electrophoresis and restriction endonuclease digestion are used to assess the integrity of the DNA. The target band should be consistent with the expected size and have a high degree of brightness concentration.

4. Sequence consistency test

The sequence information of the synthetic gene is obtained through DNA sequencing technology and compared with the set sequence to ensure a complete match. Common sequencing methods include Sanger-Couson method and Maxam-Gilbert method.