UltraBio™血液RNA稳定保存液

UltraBio™ RNA Stabilization Reagent for Blood

R751648-100ml

100ml

2-8°C储存

冰袋运输

室温或4℃保存，两年有效。

阿拉丁的UltraBio™血液RNA稳定保存液(UltraBio™ RNA Stabilization Reagent for Blood)，简称血液UltraBio™或血液UltraBio™试剂，是一种用于采集血液样品时，仅需室温操作，即可迅速抑制RNase并稳定保存血液样品中RNA的无毒无害液态试剂。本产品广泛用于新鲜血液样品中RNA的稳定保存。本产品的使用效果和液氮冻存的效果一致。本产品可以迅速渗透到血液细胞内部，完全抑制RNase，避免RNA降解，保持血液样品RNA的完整性，从而完美保存血液细胞的基因表达谱和细胞外的RNA谱，并且和液氮一样可以长期保存样品中的RNA。本产品安全便捷。本产品无任何毒性和刺激性，可室温使用，确保了使用时的安全和便捷。和使用液氮相比，可以有效避免因使用超低温的液氮而可能引起的冻伤、离心管爆裂等安全隐患，并且携带、运输非常方便。本产品可稳定血液样品的RNA表达谱。常见的血液样品保存方式是4℃短时间保存，但此时血液中细胞的生命活动还在进行，基因表达谱还会发生变化，RNA提取时与血液样品收集时的基因表达谱很可能已经出现显著差异，这种显著的差异很可能会导致实验结果的不确定性。而使用血液UltraBio™试剂保存后，血液中细胞的生命活动几乎立即被终止，RNA谱也被固定，且RNA完整性更佳(图1B)。经测试，直接将全血置于 使用说明 ： 1.对于全血样品。a.全血的收集：收集100-300µl新鲜的血液样品，加入含抗凝剂的抗凝管中，颠倒混匀。抗凝剂推荐使用EDTA钠盐或钾盐，肝素和柠檬酸盐也可以。b.保存：按照1:5的比例加入血液RNALater™试剂，例如200µl的血液加入1000µl的血液RNALater™试剂，颠倒混匀。此时血液样品在室温可以保存一周，4℃可以保存一个月，4℃保存(过夜或更长时间)后转移至-20℃或-80℃可以至少保存6个月。c.RNA抽提时样品的处理：RNA抽提前，加入等体积的DEPC水(DNase、RNase free)或其它适当的不含RNase的水，混匀，10,000×g室温离心5分钟，小心去除上清。沉淀物或剩余体积按照1:3的比例加入RNAeasy™血液RNA抽提试剂盒(离心柱式) ，如200µl的沉淀物或剩余体积加入600µl的裂解液。随后按照步骤3或RNAeasy™血液RNA抽提试剂盒(离心柱式) 使用说明进行总RNA的提取。也可以使用其它RNA抽提试剂盒中裂解液或Trizol LS等并按其说明书进行总RNA的抽提。如果希望同时抽提细胞内和细胞外的RNA，直接按照1:3的比例加入RNAeasy™血液RNA抽提试剂盒(离心柱式) 中的裂解液，例如200µl含血液RNAlater™试剂的样品加入600µl的裂解液。随后按照步骤3或RNAeasy™血液RNA抽提试剂盒(离心柱式) 使用说明进行总RNA的提取。也可以使用磁珠法血液RNA抽提试剂盒或其它RNA抽提试剂盒中裂解液或Trizol LS等并按其说明书进行总RNA的抽提；也可以使用磁珠法血液mRNA抽提试剂盒直接抽提mRNA。2.对于白细胞样品：a.白细胞的收集：取100-300µl新鲜的血液样品，加入含抗凝剂的抗凝管中，颠倒混匀。1500-2000×g，4℃离心10-15分钟。此时可见管中的液体分3层，上层为淡黄色血浆，底层为红细胞，在紧贴红细胞层上有一层灰白色的白细胞。吸掉上层的血浆，小心收集白细胞至新的离心管中，然后加入1ml PBS 洗涤，5000×g 4℃离心2分钟，小心吸除PBS，由于白细胞比较少，可以适当残留微量液体。b.保存：小心加入100-200µl的血液RNAlater™试剂，避免吹散沉淀。此时的白细胞样品在室温可以保存一周，4℃可以保存一个月，4℃保存(过夜或更长时间)后转移至-20℃或-80℃至少可以保存6个月。c.RNA抽提时样品的处理：方法1：直接按照1:3的比例加入RNAeasy™血液RNA抽提试剂盒(离心柱式) 中的裂解液，例如200µl含血液RNAlater™试剂的样品加入600µl的裂解液。随后按照步骤3或RNAeasy™血液RNA抽提试剂盒(离心柱式)使用说明进行总RNA的提取。也可以使用磁珠法血液RNA抽提试剂盒或其它RNA抽提试剂盒中裂解液或Trizol LS等并按其说明书进行总RNA的抽提。也可以使用磁珠法血液mRNA抽提试剂盒直接抽提mRNA。本方法简单，抽提获得的总RNA质量尚可，得率高，几乎没有损失，但由于没有去除血液RNAlater™试剂，纯度上可能会稍差一些。方法2：加入等体积的DEPC水(DNase、RNase free)或其它适当的不含RNase的水，10,000×g室温离心5分钟，小心去除上清，由于白细胞比较少，可以适当残留微量液体，然后加入RNAeasy™血液RNA抽提试剂盒(离心柱式) 中的裂解液300µl。随后按照步骤3或RNAeasy™血液RNA抽提试剂盒(离心柱式) 使用说明进行总RNA的提取。也可以使用磁珠法血液RNA抽提试剂盒或其它RNA抽提试剂盒中裂解液或Trizol LS等并按其说明书进行总RNA的抽提。也可以使用磁珠法血液mRNA抽提试剂盒直接抽提mRNA。本方法稍复杂，由于通过离心去除了血液RNAlater™试剂，纯度好，但在离心过程中可能会损失部分白细胞。3.血液样品总RNA的抽提：对于使用阿拉丁RNAeasy™血液RNA抽提试剂盒(离心柱式) 裂解液和抽提试剂盒的样品，建议按照以下步骤进行总RNA的抽提，或参考产品使用说明。其它RNA抽提试剂盒如磁珠法血液RNA抽提试剂盒、Trizol LS等请按其说明书进行总RNA的抽提。也可以使用磁珠法血液mRNA抽提试剂盒直接抽提mRNA。a.加入等体积结合液至裂解液中，轻轻颠倒混匀3-5次。此时可能会有沉淀物产生，属于正常现象。b.将混合物(包括沉淀物)转移至纯化柱内，12,000×g离心30秒，倒弃收集管内液体。注意：当裂解液的体积大于300µl时，在加入等体积结合液后，总体积会超过纯化柱的容量，这时应分成2次过柱，即将一半的混合物过柱后，再将剩余的混合物重复步骤b一次。c.加入600µl洗涤液I，12,000×g离心30秒，倒弃收集管内液体。d.加入600µl洗涤液II，12,000×g离心30秒，倒弃收集管内液体。e.重复步骤d一次。最高速(约14,000-16,000×g)离心2分钟，去除残留的液体。f.将RNA纯化柱置于本试剂盒提供的RNA洗脱管中，加入30-50µl洗脱液，室温放置2-3分钟，最高速离心30秒，所得溶液即为纯化的RNA。用分光光度计测浓度。本试剂盒抽提得到的RNA的A260/A280通常为2.0-2.2。A260/A230通常为1.9-2.1。注意：洗脱液需要加到纯化柱柱面中央，使其被完全吸收。如需获得更高浓度的样品，可把洗脱液的体积减小至20µl，但洗脱下来的RNA量会相对减少。室温较低时，洗脱液在37℃预热片刻对得率有所帮助。此外，洗脱后的溶液再次加回到原纯化柱再离心洗脱一次，可提高得率约10-30%；或者在第一次洗脱后使用新的洗脱液再洗脱一次，会获得约为第一次洗脱量15-40%的RNA。 Aladdin's UltraBio™ RNA Stabilization Reagent for Blood, short for Blood UltraBio™ or Blood UltraBio™ Reagent, is a non-toxic reagent that can quickly inhibit RNase and stabilize RNA in blood samples at room temperature. This product is widely used for the stabilization and preservation of RNA in fresh blood samples.This product has similar RNA stabilization effect to liquid nitrogen cryopreservation. This product can rapidly penetrate into blood cells and completely inhibit RNase to prevent RNA from degradation, thus maintaining the the integrity of RNA in blood samples. It perfectly preserves the gene expression profile in blood cells as well as the extracellular RNA profile. Similar to liquid nitrogen, it can stabilize the RNA in blood samples for a long time.This product is safe and convenient to use. This product is non-toxic and non-irritating, and can be used at room temperature. Compared to liquid nitrogen, it effectively avoids frostbite, centrifuge tube rupture and other potential risks. Moreover, it is very convenient for users to carry and transport.This product can stabilize the RNA expression profile of blood samples. The commonly used method for preserving blood samples is short-term storage at 4℃. However, at this temperature, the intracellular activities in blood cells are still active and the gene expression profile can change significantly, which causes the uncertainty in experimental results. Instead, the Blood UltraBio™ Reagent can immediately stop the intracellular activities of blood cells, thus fix the RNA profile with good RNA integrity as well, which is even better than the stabilizing effect of direct storage of whole blood at -80℃ (Figure 1B).This product is designed for stabilization and preservation of RNA (including small RNAs such as miRNA) in fresh blood samples, especially the samples that cannot be extracted immediately after collection, such as clinical samples and large amount of blood samples that cannot be frozen immediately.As shown in the table below, the storage time of blood samples in this product varies depending on the temperature.Storage Temperature37℃25℃4℃-20℃ or lower Storage Time1-2 days1 week1 month＞6 monthsThe extracted viral RNA can be used for RT-PCR, qPCR, Northern, Dot Blot, mRNA purification, in vitro translation, RNase protection30 assay, molecular cloning, microarray analysis, RNA-seq, and other applications requiring high-quality RNA.Please refer to Figure 1 for the preservation effect of this product on mouse blood samples. The whole blood samples preserved in this product at 4℃ for 30 days, at room temperature for 7 days, or at 37℃ for one day showed no RNA degradation, while most of RNA degraded in blood samples directly stored at 25℃ for 7 days (Figure 1A). This product provides better stabilization effect on the whole blood at 4℃. RNA of whole blood stored at -80℃ for 3 days partially degraded, while that of whole blood preserved in this product at -80℃ for 3 days showed almost no degradation.Figure 1. The preservation effect of Aladdin's UltraBio™ RNA Stabilization Reagent for Blood on RNA in mouse whole blood. RNA was isolated from the whole blood preserved as indicated in the figure with the RNAeasy™ Animal RNA Isolation Kit with Spin Column , then subjected to denaturing (A) and non-denaturing electrophoresis (B). This figure is for reference only.This product can also be used for the stabilization and preservation of protein and DNA in blood samples, but protein will be denatured. Therefore, the preserved samples can used for Western blot and SDS-PAGE analysis, but not enzyme activity assays.We recommend using 1ml of this product per 200µl of blood. Precautions ： This product is suitable for the preservation of fresh blood. It should be added immediately after blood collection.Samples that require long-term storage at -20℃ or -80℃ should be first stored at 4℃ overnight or longer period after the addition of this product to ensure the penetration of the Blood RNALater™ Reagent into blood cells.When thawing samples that are preserved in this product at -20℃ or -80℃, it is normal to see crystallization of the Blood RNALater™ reagent. The sample can be used for RNA extraction after the crystals are completely dissolved.All home-made reagents and consumables should be RNase-free. To prevent RNase contamination, immerse the consumables in 0.01% DEPC water overnight, followed by autoclaving and drying. Wear a disposable mask during the operation.We recommend using RNase and DNase Away to remove RNase from the working environment during the RNA extraction.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.For whole blood samples.a. Whole blood collection: Collect 100-300µl of fresh blood to a tube containing anticoagulant and mix well by inverting. We recommend using sodium or potassium EDTA as the anticoagulant. Heparin or citrate anticoagulant is also acceptable.b. Storage: Add the Blood RNALater™ Reagent into the blood sample at 5:1. For example, add 1000µl of Blood RNALater™ Reagent into 200µl of blood, and mix well by inverting. Then samples can be stored at room temperature for up to one week, or at 4℃ for up to one month. For long-term storage, place samples at 4℃ overnight or longer before transferring to -20℃ or -80℃ under which samples can be stored for at least 6 months.c. Before RNA extraction, add equal volume of DEPC water (nuclease-free) or other appropriate RNase-free water into samples and mix well, centrifuge at 10,000×g at room temperature for 5 minutes, and carefully discard the supernatant. Add Lysis Buffer provided in the RNAeasy™ Animal RNA Isolation Kit with Spin Column ( Animal RNA Isolation Kit with Spin Column at a ratio of 3:1 without centrifugation. For example, add 600µl of Lysis Buffer into 200µl of sample containing Blood RNAlater™ Reagent. Then proceed to step 3 or follow the instructions of the RNAeasy™ Animal RNA Isolation Kit with Spin Column to complete RNA extraction. 2. For leukocyte samples.a. Leukocyte collection: Collect 100-300µl of fresh blood to a tube containing anticoagulant and mix well by inverting. Centrifuge at 1500-2000×g at 4℃ for 10-15 minutes. After centrifugation, sample will display three layers, the yellow upper layer of plasma, the bottom layer of red blood cells, and the grayish white layer of leukocyte in between. Remove the upper layer of plasma and carefully collect the leukocyte into a new centrifuge tube, then wash leukocyte with 1ml of PBS ( Reagent into leukocyte and mix cell. Leukocyte in this product can be stored at room temperature for up to one week, or 4℃ for up to one month. For long-term storage, place samples at 4℃ overnight or longer before transferring to -20℃ or -80℃ under which samples can be stored for at least 6 months.c. Sample preparation before RNA extraction.Method 1: Add the Lysis Buffer provided in RNAeasy™ Animal RNA Isolation Kit with Spin Column ( Reagent containing leukocyte at the ratio of 3:1. For example, add 600µl of Lysis Buffer into 200µl of Blood RNAlater™ Reagent containing leukocyte. Then proceed to step 3 or follow the instructions of the RNAeasy™ Animal RNA Isolation Kit with Spin Column to complete RNA extraction. The lysis buffer of other RNA extraction kits or Trizol LS can also be used. This method is simple, with acceptable RNA quality and high RNA yield obtained. However, the RNA purity may be not very high due to the presence of the Blood RNAlater™ Reagent.Method 2: Add equal volume of DEPC water (nuclease-free) or other appropriate RNase-free water, centrifuge at 10,000×g at room temperature for 5 minutes, and carefully remove the supernatant. The amount of leukocyte is relatively small, so it is acceptable not to discard supernatant completely. Add 300µl of Lysis Buffer provided in the RNAeasy™ Animal RNA Isolation Kit with Spin Column ( Animal RNA Isolation Kit with Spin Column to complete RNA extraction. The lysis buffer of other RNA extraction kits or Trizol LS can also be used. This method is a little more complicated. The purity of RNA isolated by this method is increased, but some leukocyte may be lost during centrifugation.3. Extraction of total RNA from blood samples.When using the RNAeasy™ Animal RNA Isolation Kit with Spin Column ( Animal Long RNA Isolation Kit with Spin Column12TR0026RNAeasy™ Animal Long RNA Isolation Kit with Spin Column50TR0027RNAeasy™ Animal Long RNA Isolation Kit with Spin Column200TR0028RNAeasy™ Small RNA Isolation Kit with Spin Column50TR0032RNAeasy™ Animal Total RNA Isolation Kit with Spin Column50TR0046-250mlLysis Buffer for RNAeasy™ Animal RNA Isolation Kit250mlR0046-50mlLysis Buffer for RNAeasy™ Animal RNA Isolation Kit50mlR0116-100mlRNALater™ RNA Stabilization Reagent for Blood100mlR0116-500mlRNALater™ RNA Stabilization Reagent for Blood500mlR0118-100mlRNALater™ RNA Stabilization Reagent for Animal Tissue100mlR0118-500mlRNALater™ RNA Stabilization Reagent for Animal Tissue500mlR0121-100mlAllProtect™ Nucleic Acid and Protein Stabilization Reagent for Animal Tissue100mlR0121-25mlAllProtect™ Nucleic Acid and Protein Stabilization Reagent for Animal Tissue25mlR0123RNase and DNase Away250mlR0125RNase, DNase and DNA Away250mlR0141-100mlRNALater™ Viral RNA Stable Preservation Solution100mlR0141-500mlRNALater™ Viral RNA Stable Preservation Solution500ml