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View additional product information for ChargeSwitch™-Pro PCR Clean-Up Kit - FAQs (CS32050, CS32250)
9 product FAQs found
我们强烈建议使用试剂盒提供的洗脱缓冲液，并且不建议用水进行洗脱。如果你需要使用任何其它的缓冲液进行洗脱，请确保使用pH 8.5–9.0的缓冲液以实现DNA的高效洗脱。
带filter的试剂盒是基于一个双巢式柱的设计，它能够有助于通过最快最简便的步骤获得高质量的质粒（ (<10 EU/µg DNA）。经过改良的缓冲液1可去除更多内毒素。
通常而言ChargeSwitch Pro mini试剂盒从5mL菌液可得到25 µg质粒，midi试剂盒从25 mL菌液可得到200µg质粒，而 maxi试剂盒从100 mL菌液可得到800 µg质粒。试剂盒可以纯化的质粒大小范围通常是3–9 kb。
ChargeSwitch包被的磁珠是惰性的，不会影响PCR使用。我们已经发现，多达10 µL磁珠不会对常规PCR实验产生负影响。超过~10 µL的过量磁珠会抑制PCR反应。一些特殊应用可能会受到磁珠影响，包括实时定量PCR反应或MALDI-TOF。在这些应用中，应重复实验方案中描述的最终洗脱结合步骤。
We strongly recommend using the Elution Buffer provided in the kit, and do not recommend elution with water. If you need to elute in any other buffer, be sure to use a buffer of pH 8.5-9.0 for efficient DNA elution.
The filter kit is based upon a dual nested column design, which facilitates the fastest and simplest protocol for obtaining high quality plasmid (less than 10 EU/µg DNA). Modified wash buffer 1 removes more endotoxin.
The typical yield is 25 µg from a 5 mL culture for the ChargeSwitch Pro mini kit, 200 µg from a 25 mL culture for the midi kit, and 800 µg from a 100 mL culture from the maxi kit. The plasmid size range for the kits is typically 3-9 kb.
ChargeSwitch coated beads are inert and will not affect PCR. We have found that volumes of as much as 10 µL of beads have no deleterious effects on regular PCR when spiked in. Excessive quantities greater than 10 µL of beads will start to inhibit PCR. Some specific applications may be affected by beads, including real-time reactions or MALDI-TOF. In these applications, repeat the final elution binding as described in the protocol.
Based on a patented switchable chemistry, ChargeSwitch purification utilizes the charge on a solid surface for nucleic acid capture. At lower pH, the surface chemistry holds a positive charge and attracts nucleic acid. At higher pH, the surface chemistry is neutral and any bound nucleic acid is released.