Quick Hybridization Solution

快速杂交解决方案

Quick Hybridization Solution

Q743280-100ml

100ml

2-8°C储存

冰袋运输

室温或4℃保存，至少一年有效。

阿拉丁生产的Quick Hybridization Solution，也称Quick Hybridization Buffer，即UltraBio™快速杂交液，是一种用于Northern、Southern实验中RNA或DNA与RNA、DNA或Oligonucleotide探针快速高效杂交的溶液。本产品为DNase/RNase/Protease-free的无菌产品，可以和Biotin Northern Blot Kit 以及Biotin Northern Blot Kit (for Small RNA) 配合使用。本产品为高灵敏度快速杂交液，含有独特的杂交加速试剂(hybridization accelerator)，可以大大提高杂交速度和检测灵敏度，通常杂交2h即可达到杂交过夜的效果。本产品用于Northern检测小鼠肝脏总RNA中tRNA-Ala的效果参考图1，仅1ng的总RNA样品通过2h的杂交就可以获得和杂交过夜基本一致的良好检测信号。图1.使用阿拉丁生产的Quick Hybridization Solution 进行Northern检测时，分别杂交2h和杂交过夜的检测效果图。实验样品为小鼠肝脏组织中提取的总RNA，使用3’端生物素标记的Oligo探针检测。检测mouse-tRNA-Ala-TGC-3-1的探针为5’-TGGAGATGCCGGGGATCGAACC-Biotin-3’。实际检测效果会因为实验条件的不同而有所不同，图中效果仅供参考。Northern检测方法在1977年由斯坦福大学James Alwine、David Kemp和George Stark发明，可用于检测RNA的大小、丰度和电泳时的迁移位置等1, 2]。由于DNA Blot以其发明人Edwin Southern的名字命名为“Southern Blot”，与其类似的RNA Blot因此被命名为“Northern Blot” 1, 2]。Northern Blot包括以下主要步骤：RNA样品的分离与纯化，RNA样品的变性或非变性凝胶电泳，将凝胶中的RNA样品转印至尼龙膜或硝酸纤维素膜上，预杂交，与带有标记的特异核酸探针(DNA或RNA探针)杂交，最后经放射自显影、化学发光或显色的方法进行探针的检测1, 2]。其中基于化学发光的Northern检测，灵敏度非常高，又无需接触对于人体有伤害的放射性同位素，目前被广泛使用。Northern检测用于定性和定量检测目的RNA时，比qPCR或高通量测序要繁琐很多。Northern相对于qPCR和高通量测序，最大的优势是可以通过印迹检测确定目的RNA的长度或者说是电泳时的迁移位置，并且不受小RNA末端修饰的影响2, 3]。qPCR很难确定目的RNA的长度，高通量测序会受读长影响，以及RNA末端修饰的影响2, 3]。本产品为Northern检测提供了非常关键的快速高灵敏度的杂交液，给Northern杂交检测提供了极大的便利。本产品经DEPC (Diethyl pyrocarbonate，焦碳酸二乙酯)处理并高温高压灭菌处理，确保无DNase、RNase和Protease，即无DNA酶、RNA酶和蛋白酶。室温或4℃保存，至少一年有效。Quick Hybridization Solution在较低温度下容易有沉淀析出。如有沉淀出现，可在40-50℃水浴孵育，并在充分溶解后使用。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。 使用说明 ： 1.阿拉丁生产的 Quick Hybridization Solution，可用于尼龙膜上RNA或DNA样品与DNA、RNA或Oligonucleotide探针的杂交，也可以用于杂交前的预杂交。通常每100cm2尼龙膜进行预杂交和杂交时，需要使用10ml Quick Hybridization Solution。2.关于Northern检测的详细实验步骤，请参考阿拉丁生产的R0219 Biotin Northern Blot Kit或R0220 Biotin Northern Blot Kit (for Small RNA)。参考文献：1.Sabelli PA, Shewry PR. Methods Mol Biol. 1995;49:213-28.2.Segundo-Val IS, Sanz-Lozano CS. Methods Mol Biol. 2016;1434:29-43.3.Dave VP, Ngo TA, Pernestig AK, Tilevik D, Kant K, Nguyen T, Wolff A, Bang DD. Lab Invest. 2019 Apr;99(4):452-469. Aladdin's Quick Hybridization Solution is designed for rapid and efficient hybridization of RNA/DNA with RNA, DNA, or oligonucleotide probes in Northern and Southern blot experiments. This product is sterile and free of DNase/RNase/Protease. It can be used in conjunction with Aladdin's Biotin Northern Blot Kit and Biotin Northern Blot Kit (for Small RNA) .This product contains a unique hybridization accelerator, which greatly enhances the hybridization speed and the detection sensitivity. Usually, hybridization for only 2 hours can achieve the same effect as overnight hybridization. For example, when using this product in Northern blot assay of tRNA-Ala in total RNA from mouse (Figure 1), a good detection signal similar to overnight hybridization was obtained with only 1ng of total RNA after a 2-hour hybridization.Figure 1. Northern blot assay of tRNA-Ala in total RNA from mouse liver tissues using Aladdin's Quick Hybridization Solution . The 3'-biotinylated oligo probe (5'-TGGAGATGCCGGGGATCGAACC-Biotin-3) was used to hybridize the mouse-tRNA-Ala-TGC-3-1 for 2h and overnight, respectively. This figure is for reference only, which may vary due to different experimental conditions.The Northern assay was invented in 1977 by James Alwine, David Kemp and George Stark at Stanford University and can be used to detect the size, abundance and migration position of RNA after electrophoresis [1, 2]. Since the DNA Blot was named "Southern Blot" after its inventor Edwin Southern, the RNA Blot was named "Northern Blot" [1, 2]. Northern Blot consists of the following main steps Precautions ： Hyb™ Quick Hybridization Solution may precipitate at lower temperatures. If precipitation occurs, it can be incubated in a 40-50℃ water bath to dissolve the precipitates completely before use.Avoid RNase contamination during operation. Wear a disposable mask when using this product and minimize its exposure to air. After use, the lid should be immediately covered and tightened. 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.This product can be used for hybridization of RNA or DNA samples on nylon membranes with DNA, RNA, or Oligonucleotide probes. It can also be used for pre-hybridization. Generally, 10ml of Quick Hybridization Solution is needed for every 100cm2 of nylon membrane.2.For detailed procedures of Northern blot, please refer to the manual of Quick Hybridization Solution500mlR0251-500mlLow Stringency Wash Solution (DNase/RNase free)500mlR0253-500mlHigh Stringency Wash Solution (DNase/RNase free)500mlR0262-250mlNorthern Blot Stripping Solution (Mild)250mlR0266-250mlNorthern Blot Stripping Solution (Moderate)250mlR0268-250mlNorthern Blot Stripping Solution (Harsh)250mlA0303HRP-labeled Streptavidin0.2mlP0018F Ultrapure Water (DNase/RNase-Free, Sterile)100/500mlR0021DEPC-water (DNase、RNase free)100mlR0022DEPC-water (DNase、RNase free)500mlST004LAgarose50gST004B40% Acr-Bis (19:1, Nuclease free)100mlR0123RNase and DNase Away250mlR0125RNase, DNase and DNA Away250mlR0127RNase, DNase, DNA and RNA Away250ml