المهنية في صناعة الخرز المغناطيسي الحيوي
لماذا يعد تعديل الرقم الهيدروجيني ضروريًا في استخلاص الحمض النووي?
الرقم الهيدروجيني مهم جداً في استخلاص الحمض النووي والأسباب هي كما يلي:
1. Maintaining DNA Stability
DNA remains relatively stable in solutions with a pH greater than 5 وأقل من 9. When the pH is excessively high (على سبيل المثال, above 12) or low (على سبيل المثال, below 3), the hydrogen bonds between the DNA double strands will dissociate, leading to DNA denaturation. على سبيل المثال, Buffer II contains 0.2 M NaOH in plasmid DNA extraction, resulting in a system pH of 12. 6, which induces denaturation of both chromosomal and plasmid DNA. Consequently, it is necessary to adjust the pH back to neutrality in subsequent steps. This serves two purposes: first, it allows the denatured plasmid DNA to renature and remain stable; second, it leverages the difference in fragment size between genomic DNA and plasmid DNA (with plasmids being smaller and more likely to reanneal into dsDNA, while gDNA, being too long, cannot fully reanneal and instead becomes entangled, precipitating with SDS-protein complexes under the influence of high salt concentrations in buffer III).
2. Preventing DNA Degradation
EDTA can bind to metal ions such as Mg²⁺and Ca²⁺present in the solution. These metal ions are cofactors for DNase. EDTA inhibits the activity of DNase, thereby preventing the degradation of DNA by chelating these metal ions.
3. Optimizing Extraction Efficiency
Research has shown that the optimal pH for DNA extraction is 7.4. DNA extraction yield is maximized when buffer with pH 7. 4. The further the pH deviates from this value, the lower the DNA extraction yield. على سبيل المثال, the DNA extraction yield at pH 7.4 is 1.09, 1.03, 1.06, و 1.11 times that at pH 7.2, 7.6, 7.8, و 8.0, على التوالى.
Facilitating Nucleic Acid Binding to Magnetic Beads: An appropriate pH can promote the binding of nucleic acids to magnetic beads in magnetic bead-based DNA extraction methods. A high-salt binding buffer can neutralize the negative charges of nucleic acids, reducing the electrostatic repulsion between nucleic acids and magnetic beads, thereby facilitating the adsorption of nucleic acids onto the surface of the beads.
4. Protecting Lysozyme Activity
Lysozyme is utilized during the extraction of plasmid DNA from Gram-positive bacteria. Lysozyme is a glycoside hydrolase that can hydrolyze the β-1, 4 glycosidic bonds in peptidoglycan, the main chemical component of the cell wall of G+ البكتيريا, thereby exerting a lytic effect. The activity of lysozyme is inhibited when the pH of the buffer is less than 8. لذلك, it is essential to maintain an appropriate pH to ensure the activity of lysozyme.
5. Improving Elution Efficiency
The pH of the elution buffer significantly affects the efficiency of DNA elution. The pH of the elution buffer should be maintained between 7.0 و 8.5 to ensure that DNA is fully eluted. A pH that is too low will reduce the elution efficiency.
في ملخص, the adjustment of pH is crucial in the DNA extraction process. It not only helps to maintain the stability of DNA and prevent degradation but also optimizes extraction efficiency, protects the activity of relevant enzymes, and optimizes elution conditions.
مزود
شركة شنغهاي لينغجون للتكنولوجيا الحيوية, المحدودة.تأسست في 2016 وهي شركة متخصصة في تصنيع المواد المغناطيسية الحيوية وكواشف استخلاص الحمض النووي.
لدينا خبرة غنية في استخراج وتنقية الحمض النووي, تنقية البروتين, فصل الخلايا, التألق الكيميائي, وغيرها من المجالات التقنية.
منتجاتنا تستخدم على نطاق واسع في العديد من المجالات, مثل الفحوصات الطبية, الاختبارات الجينية, البحوث الجامعية, التكاثر الوراثي, وهكذا. نحن لا نقدم المنتجات فحسب، بل يمكننا أيضًا إجراء تصنيع المعدات الأصلية, أوديإم, وغيرها من الاحتياجات. إذا كان لديك حاجة ذات صلة, لا تتردد في الاتصال بنا علىsales01@lingjunbio.com.
