Ribonucleoside Vanadyl Complexes, RVC

氧钒核糖核苷复合物(Ribonucleoside Vanadyl Complexes, RVC)是一种常用的核糖核酸酶(ribonucleases, RNase)抑制剂，即RNA酶抑制剂，浓度为200mM，广泛用于各种RNA样品的分离纯化和检测过程中以抑制RNA降解。本产品通过一种优化的方法将四种rNTP的等摩尔混合物与氧化钒混合反应而成，可以和RNase以非共价键结合，形成稳定的复合物，从而抑制RNase活性。本产品是一种常用的RNase抑制剂，可广泛用于各种需要防止RNA降解的RNA分离、纯化和检测过程。本产品可以用于mRNA的纯化过程中，作为RNase的抑制剂；也可用于细胞裂解和蔗糖梯度法细胞组分分离过程中，以抑制RNase活性；还可以用于原位杂交中保护RNA探针，以及DNase I消化DNA时保护RNA等。本产品可以抑制绝大多数核糖核酸酶，但以下是例外：DNase I、S1 nuclease及Bacillus cereusribonuclease。由于本产品不抑制DNase I，所以当用DNase I消化去除RNA样品中的DNA时，可以采用本产品来抑制RNase。本产品不适合用于体外翻译体系。有报道称0.4mM以上浓度的RVC对PCR聚合酶有一定的抑制作用，所以本试剂不推荐直接用于反转录体系中，反转录体系中可以选用阿拉丁的另外一种RNase抑制剂。也可以通过后续提到的方法抽提去除RVC后再用于PCR反应。很多时候RNA样品中的RVC不必去除，可以直接用于后续的电泳、功能检测等。例如，使用本产品纯化获得的mRNA未经去除RVC，可以直接用于蛙卵的注射实验。如果需要去除RNA样品中的RVC，可以直接使用等体积苯酚进行抽提。如果苯酚中含有抗氧化剂8-羟基喹啉(8-hydroxyquinoline)，抽提含有RVC的RNA样品后会发现苯酚的颜色会变黑。当苯酚抽提样品2-4次后，苯酚的颜色不再明显变色，提示RVC已经被充分去除了。也可以在RNA沉淀前，加入RVC的摩尔数10倍量的EDTA以充分分解和失活RVC，这样可以在RNA沉淀时去除已经分解和失活的RVC。RVC浓度越高，对于RNase的抑制效果越强。参考图1，20mM的RVC可以完全抑制0.00025U/ml的RNase A。本产品浓度为200mM，一般细胞裂解液中推荐使用终浓度20mM，也可以根据经验或文献选择不同的浓度。每一批的本产品都经过RNase活性抑制的检测，检测结果参考图1。 注意事项 ： 第一次使用时建议分装，避免反复冻融。去垢剂SDS以及螯合剂EDTA可以抑制RVC的活性，请勿和RVC同时使用。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。相关产品： '], Ribonucleoside Vanadyl Complexes (RVC) is a commonly used inhibitor of various ribonucleases. The 200 mM stock solution can be widely used in isolation, purification, and assays of RNA to inhibit RNA degradation.This product is produced by reacting equimolar amounts of four rNTPs with vanadium oxide using an optimized method. It inhibits RNase activity by binding RNase to form a stable complex through non-covlent bonds.This product can be used as an RNase inhibitor in mRNA purification, cell lysis and cell fractionation by sucrose gradient method, in situ hybridization of RNA probes, and DNA digestion in RNA samples, to protect RNA from degradation.This product can inhibit most ribonucleases except DNase I, S1 nuclease and Bacillus cereus ribonuclease. As this product does not inhibit DNase I, it can be used to inhibit RNase when removing DNA from RNA samples by DNase I.This product is not suitable for use in in vitro translation systems.It has been reported that RVC at concentrations above 0.4mM has a certain inhibitory effect on PCR polymerase. Therefore, this reagent is not recommended for direct use in reverse transcription. Instead, the RNase inhibitor ( aladdin, R0102) can be used in reverse transcription. After the RVC is removed from the reverse transcription reaction by the extraction method mentioned below, PCR reactions can be performed subsequently.In many cases, RNA samples containing RVC can be used directly for electrophoresis and functional assays, etc, without the need of RNA removal. For example, mRNA containing RVC can be directly used for frog egg injection experiments .To remove the RVC from RNA samples, an equal volume of phenol can be added in the RNA sample to extract the RVC. If the phenol contains the antioxidant 8-hydroxyquinoline, the color of the phenol will turn black after the extraction of RVC from RNA samples. After 2-4 times of extraction, the color of phenol no longer changes significantly, suggesting that RVC has been thoroughly removed. Alternatively, ten times the molar amount of EDTA to RVC can be added to fully decompose and inactivate RVC before RNA precipitation, so that the decomposed and inactivated RVC can be removed during RNA precipitation.The higher the concentration of RVC, the stronger the inhibition effect on RNase. As shown in Figure 1, RNase A at 0.00025U/ml can be completely inhibited by 20mM of RVC. This product is provided in 200mM stock solution and can be diluted to a final working concentration of 20mM for general use. However, its optimal working concentration should be determined for specific experiments based on experience or literature.Figure 1. Inhibition of RNase A by different concentrations of RVC . Total RNA was incubated for 20 min with 0.00025U/ml RNase A and different concentrations of RVC as indicated, then subjected to electrophoresis analysis. This figure is for reference only, which may vary due to different experimental conditions. Precautions ： It is recommended to store this product in aliquots to void repeated freeze-thaw cycles.SDS and EDTA that can inhibit RVC activity should not be used together with RVC.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.

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