GC-rich PCR Buffer (4种套装)

GC-rich PCR Buffer

G746243-4×0.5ml

4×0.5ml

-20°C储存

超低温冰袋运输

-20℃保存。

GC-rich PCR Buffer (4管套装)提供了4种不同的GC-rich PCR Buffer，为扩增高GC含量的DNA片段提供了多种不同的选择，大大提高了高GC含量DNA片段的扩增成功率。各种GC-rich PCR Buffer可以单独使用，也可以组合使用。这样可以在更多条件下优化PCR条件。提供了推荐的几种经过优化的GC-rich PCR Buffer的组合方式，便于用户直接选用。根据相关文献报道，单一的GC-rich PCR Buffer对于某一个高GC含量的DNA片段的扩增效果很好，但对于另外一个高GC含量的DNA片段则很可能会扩增效果不太理想。正因为很难采用一种通用的GC-rich PCR Buffer 完成对于不同的高GC含量DNA片段的扩增，因此对于高GC含量DNA片段的扩增一直是分子生物学操作过程中的一个难点。针对这一实际情况，阿拉丁推出了可以给予用户多种条件进行优化组合的4管套装GC-rich PCR Buffer。本GC-rich PCR Buffer (4种套装)可以适用于各种常见的耐热DNA 聚合酶例如Taq DNA polymerase、Pfu DNApolymerase等。 注意事项 ： 在使用本试剂盒提供的GC-richPCRBuffer的同时，必须同时使用和DNA聚合酶配套的最终浓度为1X的含镁离子PCR Buffer。由于PCR反应非常灵敏可以扩增目的基因序列超过1000万倍，在操作时请注意避免微量待扩增DNA的污染，并尽量考虑设置不加模板的空白对照以确认是否有待扩增DNA的污染。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明 ： 1.PCR反应参数的设置也可以参考特定的耐热DNA聚合酶的说明进行。通常对于扩增高GC含量DNA片段的情况，延伸时间可以比常规的延伸时间适当延长一些。2.PCR反应体系的设置可以参考特定的耐热DNA聚合酶进行，仅仅添加适量本试剂盒所提供的GC-rich PCR buffer即可。可以参考如下反应体系设置PCR反应：试剂最终浓度体积体积双蒸水或Milli-Q水-(26.75-x)μl(10.7-y)μl10X PCR Buffer (with Mg2+)1X5μl2μl5X GC-rich Buffer1X10μl4μldNTP (2.5mM each)0.2mM each4μl1.6μl模板DNA10pg-1μg*xμlyμl引物混合物(10μM each)0.8μM4μl1.6μlTaq DNA Polymerase (5U/μl)1.25U/50μl0.25μl0.1μl总体积-50μl20μl3.也可以尝试采用0.5X GC-rich Buffer:试剂最终浓度体积体积双蒸水或Milli-Q水-(31.75-x)μl(12.7-y)μl10X PCR Buffer (with Mg2+)1X5μl2μl5X GC-rich Buffer0.5X5μl2μldNTP (2.5mM each)0.2mM each4μl1.6μl模板DNA10pg-1μg*xμlyμl引物混合物(10μM each)0.8μM4μl1.6μlTaq DNA Polymerase (5U/μl)1.25U/50μl0.25μl0.1μl总体积-50μl20μl4.还可以尝试在PCR反应体系中同时含有2-4种不同的1X GC-rich Buffer或0.5X GC-rich Buffer。以形成不同的组合用于优化PCR反应条件。优先推荐的GC-rich Buffer 及其组合(最终浓度)包括：a. 1X GC-rich Buffer Ab. 1X GC-rich Buffer Bc. 0.5X GC-rich Buffer Bd. 1X GC-rich Buffer C e. 2X GC-rich Buffer Cf. 1X GC-rich Buffer C + 1X GC-rich Buffer Dg. 2X GC-rich Buffer C + 1X GC-rich Buffer Dh. 1X GC-rich Buffer C + 0.2X GC-rich Buffer B i. 2X GC-rich Buffer C + 0.2X GC-rich Buffer B 5.可以根据初步的PCR扩增效果，再对效果较好的GC-rich Buffer再进行优化组合，进一步优化GC-rich Buffer的组合。最后根据PCR反应的扩增效果，选择最佳的GC-rich PCR buffer或其组合。 GC-rich PCR Buffer Set provides 4 GC-rich PCR Buffers that can be used individually or conjunctly to improve the amplification of DNA fragments with high GC content in PCR.According to relevant literature, it is difficult to use a universal GC-rich PCR Buffer to improve the amplification of all DNA fragments with high GC content. Therefore, aladdin developed this GC-rich PCR Buffer Set for users to optimize the PCR conditions conveniently. Several recommended combinations of GC-rich PCR Buffers are also provided for users to use directly.This product is compatible with a variety of commonly used heat-resistant DNA polymerases, such as Taq DNA polymerase and Pfu DNA polymerase. Precautions ： The GC-rich PCR Buffer provided by this kit must be used in conjunction with the magnesium-containing PCR Buffer for a particular DNA polymerase.As PCR reaction is extremely sensitive, any contamination of DNA samples should be avoided. Always set up blank controls without templates.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use ： 1. The PCR reaction can be carried out by referring to the instructions of the DNA polymerase to be used. Generally, the extension time can be appropriately extended to amplify DNA fragments with high GC content. 2. Set up the PCR reaction in a DNase-free microfuge tube as follows:ComponentFinal ConcentrationVolumeVolumeddH2O or Milli-Q water-(26.75-x)µl(10.7-y)µl10X PCR Buffer (with Mg2+)1X5µl2µl5X GC-rich Buffer1X10µl4µldNTP (2.5mM each)0.2mM each4µl1.6µlTemplate DNA10pg-1µg*xµlyµlPCR Primer Mix (10µM each)0.8µM4µl1.6µlTaq DNA Polymerase (5U/µl)1.25U/50µl0.25µl0.1µlTotal Volume-50µl20µlOr:ComponentFinal ConcentrationVolumeVolumeddH2O or Milli-Q water-(31.75-x)µl(12.7-y)µl10X PCR Buffer (with Mg2+)1X5µl2µl5X GC-rich Buffer0.5X5µl2µldNTP (2.5mM each)0.2mM each4µl1.6µlTemplate DNA10pg-1µg*xµlyµlPCR Primer Mix (10µM each)0.8µM4µl1.6µlTaq DNA Polymerase (5U/µl)1.25U/50µl0.25µl0.1µlTotal Volume-50µl20µl3. More than one GC-rich Buffers can be added in one PCR reaction mix. The preferred GC-rich Buffers and their combinations in a PCR reaction mix are listed as follows:a. 1X GC-rich Buffer Ab. 1X GC-rich Buffer Bc. 0.5X GC-rich Buffer Bd. 1X GC-rich Buffer C e. 2X GC-rich Buffer Cf. 1X GC-rich Buffer C + 1X GC-rich Buffer Dg. 2X GC-rich Buffer C + 1X GC-rich Buffer Dh. 1X GC-rich Buffer C + 0.2X GC-rich Buffer B i. 2X GC-rich Buffer C + 0.2X GC-rich Buffer B 4. According to the preliminary amplification effect, the combination of GC-rich Buffers can be further optimized to determine the best GC-rich PCR Buffer or combination for the amplification of target DNA.