1、 48 gene synthesizer equipment related errors:

1. Sampling system error

Fluctuations in pipetting accuracy:

There may be differences in the pipettes or valves of different channels, resulting in inconsistent sample volumes (such as uneven distribution of base monomers).

After long-term use, the pipetting components may wear or become clogged, resulting in volume deviation.

Cross contamination:

Insufficient sealing between channels can lead to liquid residue or volatile diffusion, causing base or enzyme contamination.

2. Temperature control error

Reaction system temperature difference:

The temperature uniformity of the 48 channel heating module is insufficient, and some hole positions have high or low temperatures, which affects the extension efficiency.

Inconsistent heating/cooling rates result in asynchronous PCR or ligation reactions.

Uneven hot cap pressure:

The contact between the hot cover and the reaction plate is not tight, and some of the pores evaporate severely, changing the volume of the reaction system.

3. Detecting system errors

Fluorescence/absorbance detection deviation:

The sensitivity difference of optical detectors in different channels leads to misjudgment of synthesis efficiency.

Background noise interference (such as fluorescence quenching or scattering) affects the accuracy of real-time monitoring.

4. Mechanical failure

Uneven separation or oscillation of magnetic beads:

The difference in magnetic field strength of the magnetic module leads to different magnetic bead recovery efficiency, which affects the washing or purification effect.

Inconsistent oscillation frequency or time leads to insufficient reaction mixing.

2、 Operation and process errors of 48 gene synthesizer:

1. Program setting error

Parameter input deviation:

Improper setting of synthesis cycles (such as extension time and denaturation temperature) can lead to an increase in fragment length or mismatch rate.

Parameters were not adjusted based on template types such as high GC content and secondary structure.

Automated scripting vulnerability:

Improper connection of multi-step programs (such as cutting, connecting, purifying) can lead to reaction interruption or excessive processing.

2. Sample loading issue

Sample addition order or position error:

Failure to follow standard procedures during manual sample addition (such as failure to replace the nozzle) resulted in the introduction of exogenous DNA contamination.

The difference in the amount of starting materials in different channels (such as different primer concentrations) leads to uneven synthesis efficiency.

3. Low purification and recovery efficiency

Purification loss of magnetic beads or silica gel membrane:

The adsorption loss of small fragments of DNA (such as short oligonucleotides) during the purification process reduces yield.

Incomplete washing, residual salt or impurities inhibit subsequent reactions.