Highlights
- Easy Handling: Bypass chloroform, phase separation and precipitation steps.
- NGS-Ready: Ultra-pure RNA without phenol carryover. No DNA contamination (DNase I included).
- Non-Biased: Complete RNA recovery without miRNA loss.
Description
| Compatibility | TRIzol®, RNAzol®, QIAzol®, TriPure™, TriSure™ and all other acid-guanidinium-phenol based solutions can be used in place of TRI Reagent®. |
|---|---|
| Equipment | Microcentrifuge, vortex, magstand |
| Sample Inactivation | TRI Reagent® (provided with R2101, R2103, and R2105) inhibits RNase activity and inactivates viruses and other infectious agents. |
| Sample Source | Any sample stored and preserved in TRI Reagent®, TRIzol® or similar (animal cells, tissue, bacteria, yeast, fecal, biological fluids, and in vitro processed RNA (e.g., transcription products, DNase-treated or labeled RNA)). |
| Size Range | Total RNA ≥ 17 nt |
| Yield | 10 µg RNA (binding capacity), ≥ 30 µl (elution volume) |
Q1: Is Direct-zol suitable for very small numbers of cells?
Yes, the Direct-zol MicroPrep (#R2060) is designed and capable of purifying RNA down to single cell inputs (picogram amounts). A sensitive quantification method is needed (e.g. Qubit, qPCR, etc.)
Q2: Is DNase I available for individual purchase?
All kit components are available for purchase separately.
Q3: How to store DNase-I following resuspension?
Lyophilized DNase I is stable at room temperature. Once resuspended, store frozen aliquots. Minimize freeze thaw cycles as much as possible. Freeze thaw will lower DNase activity.
Q4: Is the DNase-I treatment necessary?
If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.
Q5: Is the kit compatible with samples stored in DNA/RNA Shield?
Yes, bring samples homogenized and stored in DNA/RNA Shield to room temperature (20-30ºC). Add 3 volume of TRIzol/TRI Reagent and mix well. Proceed with RNA Purification.
Q6: Is it possible to extract proteins with the Direct-zol RNA kits?
Yes, proteins can be Acetone Precipitated post RNA binding step. Please request supplementary protocol from Zymo Research Technical Support.
Q7: Can samples be stored in TRIzol/TRI Reagent prior to processing?
Yes, samples in TRIzol/TRI Reagent or similar are stable overnight at room temperature and can be stored frozen (-80C). Be sure to lyse and homogenize the sample well prior to freezing. Bring the sample to room temperature prior to RNA Purification.
Q8: Is it possible to isolate DNA with the Direct-zol RNA kits?
Direct-zol DNA/RNA (D2080) kits can isolate DNA from TRIzol
Q9: Is the RNA suitable for Next-Gen sequencing or other sensitive downstream applications?
Yes, the RNA is high quality (A260/A280 >1.8, A260/A230 >1.8) and suitable for any downstream application, including NGS, RT-PCR, hybridization, etc.
Q10: Which phenol-based reagents are compatible with Direct-zol?
The Direct-zol kits are compatible with TRI Reagent, TRIzol, Qiazol, RNAzol, TriPure, TriSure, etc., and any other acid-guanidinium phenol-based reagents.
Q11: What is the difference between the Direct-zol RNA and Quick-RNA kits?
Direct-zol is for samples stored/collected into TRIzol/similar reagents. Quick-RNA is for all other samples.
Q12: What is the difference between the Direct-zol RNA MiniPrep and the Direct-zol RNA MiniPrep Plus?
Both kits function the same, the only difference is the RNA binding capacity of the column provided with the kit.
Q13: I ran out of RNA Wash Buffer. Can I use something else?
Yes, use 80% ethanol as a substitute. RNA Wash Buffer is also sold separately.
“Before I discovered this kit, I was isolating RNA the old school way with chloroform and it would take half the day to finish the protocol. The Direct-zol RNA Miniprep kit is AWESOME!It took hardly any time, the protocol was so easy, and my RNA quality was SO much better. Honestly, this kit revolutionized my life at the bench.”
-A. Newhart (The Wistar Institute)
“Simple protocol and yielded good quality of RNA. Only one kit working for all type of tissue, cell and especially biological fluids.”
-Mohan K. (University of Illinois, Chicago)
“Previously I used a protocol that took 3 hours, now I can have my RNA in 20 minutes. What is not to like about that? Just one column and two buffers, I love it.”
-Arjan V. (Indiana University)
Read More| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| D4100-2-3 | MagBinding Beads | 3 ml | $66.00 | |
| D4100-2-12 | MagBinding Beads | 12 ml | $125.00 | |
| W1001-30 | DNase/RNase-Free Water | 30 ml | $22.00 | |
| R1060-2-100 | RNA Prep Buffer | 100 ml | $122.00 | |
| R1060-2-25 | RNA Prep Buffer | 25 ml | $40.00 | |
| R2100-1-20 | Direct-zol Binding Buffer Concentrate | 20 ml | $84.00 | |
| R2100-2-200 | Direct-zol MagBead PreWash | 200 ml | $174.00 | |
| R2130-1-120 | MagBead DNA/RNA Wash 1 | 120 ml | $198.00 | |
| R2130-1-30 | MagBead DNA/RNA Wash 1 | 30 ml | $63.00 | |
| R2130-2-20 | MagBead DNA/RNA Wash 2 | 20 ml | $54.00 | |
| R2130-2-80 | MagBead DNA/RNA Wash 2 | 80 ml | $171.00 | |
| C2002 | Collection Plate | 2 Plates | $22.00 | |
| C2003 | Elution Plate | 2 Plates | $19.00 | |
| C2007-8 | 96-Well Plate Cover Foil | 8 Foils | $18.00 | |
| R2050-1-200 | TRI Reagent | 200 ml | $219.00 | |
| E1010-1-4 | DNA Digestion Buffer | 4 mL | $15.00 | |
| E1010 | DNase I Set | 250 U | $56.00 |
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
细胞质中有各种细胞器,细胞内的化学反应大多都是在细胞质中进行,因此是代谢中心.
细胞核中有DNA,是遗传信息库,是细胞代谢和遗传的控制中心.
康乃尔大学应用暨工程物理学（AppliedEngineeringPhysics）副教授KarlA.Kasischke等人成功利用多光子显微技术发现脑部神经细胞（neurons）和星状细胞(astrocytes)之间的如何地交互作用来燃烧氧气和葡萄糖进行糖解作用(glycosis)﹐以达到脑部特别能量的需求。其结果已发表于今年七月的《科学》（Science）杂志上。
该研究团队表示﹐他们根据大脑代谢的辅?烟碱醯胺腺嘌呤双核甘酸（NADH,nicotinamideadeninedinucleotide）两种不同能源状态的影像﹐将最具争议性脑细胞能量代谢的星状细胞—神经元乳酸穿梭（theastrocyte-neuronlactateshuttle）假设作确认与再定义。
KarlA.Kasischke说道﹐在过去十年当中﹐科学家们激烈争议讨论﹐被激活的大脑究竟是进行有氧代谢把葡萄糖彻底分解成水？还是进行无氧状态的糖解作用产生乳酸（lactate）？他表示﹐他们的研究已经发现星状细胞糖解作用伴随着神经活化引发神经性氧化代谢（NeuronalOxidativeMetabolism）将这两种目前对立的说法产生一致性并造成两派双赢的局面。由于他们所使用的多光子显微镜可以让NADH产生内生性荧光影像﹐显示出脑神经内早期氧化代谢终究是持续的﹐并且在约10秒后让星状细胞—神经元乳酸穿梭（theastrocyte-neuronlactateshuttle）作脑细胞晚期的活化作用。神经细胞甚至在休息的时候是不断代谢葡萄糖﹐并且当讯号开始穿越神经细胞时﹐代谢葡萄糖的现象会持续表达﹐然后星状细胞会将代谢葡萄糖所得到的乳酸﹐提供出来做为燃料。
目前医师所使用的脑神经影像技术﹐例如功能性磁共振影像（fMRI,functionalmagneticresonanceimaging）和正子造影系统（PET；positronemissiontomography）虽然可分别探测血流和血氧变化﹐提供医师了解大脑功能变化﹐但是在时间和空间的分辨率却无法满足研究人员的需求。而相较之下﹐多光子显微技术却能提供中枢神经系统（CNS；centralnervoussystem）高分辨率﹐3D立体的组织影像﹐强力地帮助研究人员探讨脑细胞代谢途径。
这场十多年来的争论﹐看来各持己见的双方都没有输。不过﹐最重要的意义是﹐多光子显微技术足以提供大脑代谢等研究功能性方面的应用﹐并且提供给医师较佳的方式来观察中风或阿兹海默症等脑部损害。
全文链接：http://www.sciencemag.org/cgi/reprint/305/5680/50.pdf
解析：1.细胞质基质（也叫胞质溶胶）是指除细胞器外细胞质的其余部分。细胞质基质是活细胞进行新陈代谢的主要场所。
2.细胞新陈代谢的次要场所是：细胞核、线粒体基质、叶绿体基质等基质。
酶的作用是催化剂,促进或抑制反应的进行.加热主要是通过升高温度加快反应速率,无机催化剂和酶的原理相同,都是通过降低反应的活化能加快反应速率
A.衰老的细胞新陈代谢速率加快
B.在衰老的细胞内有些酶的活性降低
C.衰老的细胞呼吸速率减慢
D.细胞膜通透性改变，使物质运输功能降低
2015年7月6日讯/生物谷BIOON/--最近，来自艾默里大学的科学家发现在许多黑色素瘤中存在一个重要基因突变能够使癌细胞的代谢重新连线，使癌细胞的生长依赖于一种参与酮体生成的催化酶，这一发现为解决黑色素瘤细胞对靶向药物的抵抗，开发新的替代药物提供了深入见解，同时也部分解释了为什么这一突变在黑色素瘤细胞中频发。近日，相关研究结果发表在国际学术期刊molecularcell。
B-raf基因发生V600E突变在黑色素瘤细胞中非常常见，这一突变能够促进癌细胞生长，除了在黑色素瘤中存在，在一些结肠癌和甲状腺癌病例中也发现存在B-rafV600E突变。目前已经开发出一些针对B-rafV600E基因突变的靶向药物，但在临床实验中发现，在癌症得到明显改善之后，携带V600E基因突变的癌细胞都不可避免地产生药物抗性。
在这项研究中，研究人员发现B-rafV600E基因突变能够刺激癌细胞产生更多的HMG-CoA裂解酶，携带V600E突变的黑色素瘤细胞生长非常依赖于该酶，而其他的黑色素瘤细胞则不会。HMG-CoA裂解酶是酮体生成途径中一个重要酶，能够帮助机体在血糖水平较低时降解脂肪酸以获得能量。酮体生成途径能够受到低糖，高脂饮食刺激激活，通常发生于肝脏，但B-rafV600E基因突变启动了癌细胞中的酮体生成，以维持癌细胞生长存活。除此之外，研究人员还发现酮体生成途径的重要产物乙酰乙酸能够刺激携带B-rafV600E基因突变的癌细胞继续增殖。
总得来说，这项研究证明B-rafV600E基因突变能够使黑色素瘤细胞中的代谢途径重新连线，增强癌细胞对酮体生成途径的依赖性，这一发现对于解决黑色素瘤细胞对靶向药物的抵抗，开发新的替代药物具有重要意义。（生物谷Bioon.com）
不破坏细胞结构，同步动态侦测细胞有氧呼吸，糖酵解OCR/ECA(总或乳酸ECA)；
i.过氧敏感荧光素（或pH敏感荧光素）,Ex340-380/535/Em630-680nm，实时测量线粒体/胞内/胞外重要代谢指标，荧光素为非结构性结合可逆转改变；
ii.可同时或单个测量OCR/ECA或其他参数，不会做成浪费，可采用时间分辨荧光技术可加强系统信噪比
iii.通过两个加药口，可对检测细胞加入适当抑制剂/刺激实时监测细胞对不同毒素作用
