RNA原位杂交

Prior to Extractions ・ Bake all necessary glassware metal spatulas mortars and pestles overnight in a 200℃ oven after wrapping them in aluminum foil. For each sample you should minimally prepare one ...

Polyacrylamide Gel Electrophoresis (PAGE) for use withRibonuclease Protection Assay (RPA):1. Making the Gel: 5% Denaturing gel for Ribonuclease Protection Assay: Ureahigh quality ...

1. Wear gloves at all time including filling pipet tip in racks filling jars with Eppendorf tubes and weighing chemicals to prepare solutions.2. Pull weighing paper or weighing boat fr ...

在所有RNA实验中，最关键的因素是分离得到全长的RNA。而实验失败的主要原因是核糖核酸酶（RNA酶）的污染。由于RNA酶广泛存在而稳定，一般反应不需要辅助因子。因而RNA制剂中只要存在少量的RNA酶就会引起RNA在制备与分析过程中的降解，而所制备的RNA的纯度和完整性又可直接影响RNA分析的结果，所以RNA的制备与分析操作难度极大。在实验中，一方面要严格控制外源性RNA酶的污染；另一方面要最大限度 ...

警告: 有毒物接触皮肤或者不慎吞服。会导致灼伤。一旦接触皮肤后立即以大量的洗涤剂和清水清洗。若感不适，看医生并寻求苯酚和其他成分(JTSRN 80100437-5000p)的正确治疗方案。） TRIZOL在室温下能稳定保存12个月。尽管如此，为达到最佳效果，我们建议保存在2―8°C的环境下。一般描述：TRIZOL试剂是直接从细胞或组织中提取总RNA的试剂。它在破碎和溶解细胞时能保持RNA的完整性 ...

Use DeliverX Transfection from PanomicsModified protocol belowStep 1: Make complexesNote: Do not scale up more than 3xThaw siRNA and DeliverX reagentDilute siRNA into buffer 110 uM siRNA 2.17 ulBuf ...

Using siRNA for gene silencing is a rapidly evolving tool in molecular biology. There are several methods for preparing siRNA such as chemical synthesis in vitro transcription siRNA expression vectors ...

一、实验目的 （1）熟悉酵母RNA的提取方法（2）掌握地衣酚显色法测定酵母RNA含量的原理和方法二、实验原理 RNA与浓盐酸共热时，即发生降解，形成的核糖继而转变成糠醛，后者与3， 5-二羟甲苯反应呈鲜绿色，该反应需用三氯化铁和氯化铜作催化剂，反应产物在670nm处有最大吸收。RNA在20－250微克范围内，光吸收与RNA浓度 成正比。地衣酚反应特异性较差，凡是戊糖均有此反应。RNA和其他杂质 ...

一、实验目的（1）了解制备RNA几种方法的原理及优缺点（2）掌握稀碱法提取兔肝RNA的原理和操作技术二、实验原理 细胞内大部份RNA均与蛋白质结合在一起，并且多以核蛋白的形式存在。因此，分离制备RNA时，首先遇到的是必须使RNA与蛋白解离，并除去蛋白质。由于RNA的种类来源和存在形式不同，所用的制备方法各异，一般常用的方法有，盐酸法，稀碱法和苯酚法。其中，以苯酚法使用较为广泛。产品纯度高，适合小规 ...

在收集到生物材料之后，最好能即刻进行RNA 制备工作。若需暂时储存，则应以液氮将生物材料急速冷冻后，储存于-80℃冷冻柜。在制备RNA 时，将储存于冷冻柜的材料取出，立即以加入液氮研磨的方式打破细胞，不可以先行解冻，以避免RNase 的作用。1. 提取组织RNA 时，每50～100mg 组织用1ml Trizol 试剂对组织进行裂解；提取细胞RNA 时，先离心沉淀细胞，每5-10 �106 个细胞 ...

收集细胞，离心5000r/min ，5分钟，弃上清，加入异硫氰酸胍变性液（异硫氰酸胍4mol/L；柠檬酸钠25mmol/L，Sarkosy10.5%β-巯基乙醇0.1mol/L）500ml，混匀，振荡10秒，加2 mol/L醋酸钠50ml，水饱和酚500m l和氯仿/异戊醇（49：1）100m l颠倒混合数秒钟，冰浴15分钟，4℃离心10000 r/min，15分钟，吸取上清，加入等体积异丙醇或2 ...

IntroductionExpression profiling of cells is a valuable tool in understanding gene regulatory networks and in identifying genes necessary for normal developmental processes. In the haematopoietic syst ...

IntroductionFluorescence in situ hybridization (FISH) has become a widely used method in genome and molecular genetic studies. The technique is highly versatile and has been adapted to carry out genom ...

IntroductionThe recent discovery of RNA interference and in particular the observation that siRNAs can modulate gene expression at the level of transcription i.e. small-interfering RNA (siRNA) directe ...

IntroductionRNAse A treatment of permeabilized cells followed by immunostaining is a method which allows to show if the localization of a protein into the cell involves an RNAse-sensitive structure. T ...

Notes:RNA should be at 1 uM concentration (1 pmole/uL) There are 3 conditions (use a blank/background reaction for each condition). SolutionsCE Buffer (10X)1M Na Cacodylate (4.28 g into 20mL H2O) pH ...

MaterialsDNA: Höchst 33258 (High range solution): (Invitrogen H1398 or Sigma B1155) 1µg/mL in 1x TNE Filter and store at 4ºC in an amber bottle. 10x TNE stock: 100 mM Tris 2 M NaCl 10 mM EDTA ...

Procedure1. Homogenize cells (10 million) or tissue (50-100 mg) in 1 mL TRIzol Reagent (e.g. scrape and pass through 30G needle dounce homogenize and pass through needle or use a homogenizer) and tra ...

Materials Qiagen RNEasy Midi Kit Dounce homogenizers (for tissues) Syringes and needles (for cultured cells) 70% EtOH Sterile 15 mL conical tubes Procedure 1) Before using the reagents for the first ...

MaterialsMessageAmp II aRNA Amplication Kit (Cat #1751 Ambion)100% EthanolRNA samples (e.g. 1 µg RNA per sample)ProcedureReverse transcriptionVerify that EtOH (24 mL) has been added to the Wash Buffer ...