The extraction result may be influenced, if an un-fresh reagent is used. Use reagents after purchase without storage for an extended period of time.
The axis of the system may be disoriented during move, for example, by impact. Operation of the system in such a state may result in troubles and failure of the system. The system should be readjusted after move of the system (especially with impact). Please contact us.
Use of the apparatus, after the reagent or sample is aspirated into it, may lead to leakage and fluctuation of aspirating/dispensing amount. It affects system operation and extraction result, and thus, please contact us as soon as possible.
Deterioration in piercing efficiency for example by corrosion of the piercing pin end may lead to easier piercing defect of the aluminum seal. Use of it, as it is, may lead to failure of the system. As it will demand maintenance of the system, please contact us.
Stainless steel is used for example for reagent tank rack, and will cause rusting on it when used chlorine sanitizer during cleaning. If a chlorine sanitizer is used, make sure to wipe the surface with alcohol or water, so that no chlorine sanitizer remains on the stainless steel position.
The yield becomes smaller, when the copy number of plasmid DNAs in sample is low. Increase the amount of the microbe used. (The system is compatible with 1 ml of culture solution having an O.D.660 of up to 9.0.)
When an excessive amount of sample is used, magnetic particles become hardly dispersible in the steps of adsorption and washing process for nucleic acids and the purity of the extract may decline. Use a microbial cell suspension with an O.D.660 of 9.0 or less.
It may be because of inhibition by ethanol present in a trace amount in the extract. The extract, when used in an amount of more than 1/5 of the sequencing or PCR reaction solution, may inhibit the reaction and thus, reduce the amount of the extract used. Alternatively in the case of the inhibition by ethanol, heating of the extract (75°C, 40 minutes) may reduce the inhibition.
The extract seems brown in color, if it is contaminated with magnetic particles. Although there is no adverse influence on downstream steps, it is preferably to centrifuge the solution (10,000 g, 1 minute) for removal of the magnetic particles.
Make sure that there is no problem in the storage temperature of the blood sample use. Please use the sample in extraction operation, after it is heated up sufficiently to room temperature. In the case of a blood sample stored refrigerated, the yield may become smaller when the storage period is elongated. Make sure that there is no problem in storage temperature and place of the reagents. Storage of reagents in a place under vibration may lead to deterioration in performance of the magnetic particles. Avoid storage in a place under excessive vibration.
When Tris-HCl is selected as eluate, there may be some influence, depending on the PCR reaction condition. In such a case, use the sample after extraction operation carried out by using distilled water as the eluate.
There are many possible causes.
(1) Excessive amount of sample
Use of a sample in an amount more than a particular value does not lead to increase, but leads to decrease of the yield. Reduce the sample amount. In the case of a cell sample, use it at a concentration of 1×106 cells/200 μl or less, while in the case of a tissue sample, use it at a concentration of 10 mg or less/150 μl.
(2) Insufficient solubilization and homogenization of sample
Agitation or homogenization of the sample may be insufficient. Extraction treatment in the state where solid matters remain in the solution because of insufficient homogenization may cause tip clogging during extraction operation, leading to decrease of the yield.
(3) Tip clogging
Some tissues contain many muscle fibers and thus, make sure to conduct homogenization operation sufficiently.
(4) Condition of prepacked reagents
Tap or shake the reagent cartridge gently, when the cartridge reagent contains air bubbles or when there is deposition of reagents or water droplets in the reagent-sealing region or in the upper region of internal cartridge well. Processing with residual air bubbles may prohibit complete aspirating of the reagent and cause, for example, foaming during agitation, which often results in deterioration of the yield.
The purity may decline, when an excessive amount of sample is used. Reduce the sample amount before extraction operation (click here to see relevant Q&A/FAQ). When the RNA concentration in the extract is low, the A260/A280 value may decline. Contamination of magnetic particles in the extract may lead to increase of background (A320) or generation of noises. Use the supernatant after centrifugation (10,000 g, 1 minute) in absorbance measurement and various electrophoretic analyses.
The RNases may not be inactivated sufficiently, if an excessive amount of sample is used. Reduce the sample amount before extraction operation (click here to see relevant Q&A/FAQ). Sample-collecting operation, storage state and homogenization-solubilization condition also have an influence on decomposition of RNAs, and close attention should be given to these parameters as well. In addition, avoid storage of the extract for a long period and store it soon in a deep freezer (-80°C).
If there are intron-less genes and genes very similar in sequence on genome, influence by the genome DNA should be taken into consideration. Use a total RNA extracted by a protocol including DNase treatment. When an excessive amount of sample is used, it may influence on the extraction yield, purity and decomposition of total RNA (see Q&A/FAQ: sample quantity, RNA purity and RNA decomposition).
Extraction reagents and sample impurities may remain in and exert an influence on the next steps, depending on the state of the sample (storage period, storage temperature, etc.). Please confirm that the desired genome DNA is extracted without any problem by electrophoresis or absorbance measurement.
The system is reset to the initial state during resumption after power failure. It is thus impossible to resume continuous operation. Would you please set samples, consumables and others and start operation from the first once again.
Make sure to confirm that the AC power source cable is connected to the plug socket and the apparatus reliably. Use the AC power cable attached. If there is no power supply even when the AC power cable is connected, the fuse of the apparatus may be disconnected. Contact us in such a case.
The number of the samples cannot be increased during processing. Please do not open the door or place your hand or consumables into the apparatus during operation, because it may result in failure of the apparatus and injury of the operator.