बायोमैग्नेटिक मोतियों के पेशेवर निर्माता
पीसीआर पर चुंबकीय मोतियों के निरोधात्मक प्रभाव को कैसे कम किया जा सकता है?
With the advancement of
precision medicine and the reduction in NGS sequencing costs,magnetic bead-based nucleic acid extraction has become standard practice in molecular diagnostics laboratories. To achieve higher recovery rates and avoid errors caused by incomplete elution, an increasing number of researchers opt to add magnetic beads capturing nucleic acids directly into the PCR system for amplification.
तथापि
While this “PCR with Magnetic beads” approach streamlines workflows, it has introduced insidious frustrations for many practitioners: PCR amplification failures, elevated Ct values, or even absent amplification curves. The culprit behind these issues is often the magnetic beads themselves, which inhibit the PCR system.
Many intuitively assume magnetic beads are merely inert solid carriers that should not interfere with amplification. Yet experimental data consistently demonstrates widespread bead-induced PCR inhibition.
Preliminary experiments employing our magnetic bead products 160011, 160013, और 220015 for bead-in-PCR revealed:
| नहीं. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
| MB | 160011（50mg/ml） | 160013（50mg/ml） | 220015（50mg/ml） | ||||||
| Content | 0.5एमजी | 0.75एमजी | 1एमजी | 0.5एमजी | 0.75एमजी | 1एमजी | 0.5एमजी | 0.75एमजी | 1एमजी |
Result as follow：
Based on electrophoresis results, amplification efficiency at a magnetic bead concentration of 0.5 एमजी: 220015 < 160013 < 160011; Given the known magnetic bead particle sizes: 220015 < 160013 < 160011, and considering their specific surface areas: 160011 < 160013 < 220015, the binding sites per bead under identical conditions are thus: 160011＜160013＜220015; This may explain why PCR inhibition becomes more pronounced as magnetic bead concentration increases.
फलस्वरूप
A special component—an enhancer—is required in this PCR system. Common enhancers include betaine, trehalose, DMSO (dimethyl sulfoxide), glycerol, and BSA (bovine serum albumin). Among these, BSA acts as a sacrificial substrate, preferentially binding to magnetic beads to shield the polymerase from inhibitor interference.
So we sought to determine whether reducing magnetic bead concentration (0.025 mg–0.075 mg) supplemented with 100 μg/ml BSA as a PCR enhancer would yield more favourable results.
| नहीं. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
| MB | 160011（50mg/ml） | 160013（50mg/ml） | 220015（10mg/ml） | ||||||
| Content | 0.025एमजी | 0.05एमजी | 0.075एमजी | 0.025एमजी | 0.05एमजी | 0.075एमजी | 0.005एमजी | 0.01एमजी | 0.015एमजी |
Result as follow：
It can be observed that at 0.025 एमजी, 160011 और 160013 exhibit no difference from the control, whereas at 0.05 mg and 0.075 का, their efficacy diminishes progressively. 220015, consistent with previous analyses, not only exhibits excellent suspension properties but also possesses a large specific surface area with numerous binding sites. This enables effective adsorption of Taq polymerase, thereby inhibiting amplification. When the magnetic bead concentration was reduced to 0.005 एमजी, the band intensity reached only half that of the control.
Following the foundational optimisation under these conditions, our 220013 product shall be selected for efficacy validation.：
| नहीं. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| MB | 220013（10mg/ml） | 220015（10mg/ml） | ||||||
| Content | 0.003एमजी | 0.005एमजी | 0.007एमजी | 0.01एमजी | 0.003एमजी | 0.005एमजी | 0.007एमजी | 0.01एमजी |
Result as follow：
It can be observed that the performance outcomes of 220013 और 22015 are indistinguishable.
The aforementioned magnetic beads 160011, 160013, 220013, और 220015 can all achieve optimal results in PCR through system optimisation. They are suitable for application in POCT rapid testing, pathogenic microorganism detection, automated extraction, and similar scenarios.
देने वाला
शंघाई लिंगजुन बायोटेक्नोलॉजी कंपनी, लिमिटेडमें स्थापित किया गया था 2016 जो बायोमैग्नेटिक सामग्रियों और न्यूक्लिक एसिड निष्कर्षण अभिकर्मकों का एक पेशेवर निर्माता है.
हमारे पास न्यूक्लिक एसिड निष्कर्षण और शुद्धिकरण में समृद्ध अनुभव है, प्रोटीन शुद्धि, कोशिका पृथक्करण, chemiluminescence, और अन्य तकनीकी क्षेत्र.
हमारे उत्पाद कई क्षेत्रों में व्यापक रूप से उपयोग किए जाते हैं, जैसे चिकित्सा परीक्षण, आनुवंशिक परीक्षण, विश्वविद्यालय अनुसंधान, आनुवंशिक प्रजनन, और इसी तरह. हम न केवल उत्पाद उपलब्ध कराते हैं बल्कि ओईएम का कार्य भी कर सकते हैं, ओडीएम, और अन्य जरूरतें. यदि आपको इससे संबंधित कोई आवश्यकता है, हमसे संपर्क करने के लिए कृपया स्वतंत्र महसूस करें .
