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®. |
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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 MoreCat # | Name | Size | Price | |
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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 |
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我可以理解为,
丙氨酸+α-酮戊二酸→转氨酶→谷氨酸+丙酮酸
一个意思吗?
书上没有相关说法,求解
有机学的不好,各位大神,谁知道酮羰基为什么比酯基和酰氨基稳定吗?感谢!!!!!
生理的问题:α酮戊二酸是怎么生成两分子碳酸氢根的?
“不能让它就这么停了,太可惜了!”作为临床医药出身的王杰深知“美她司酮”身上闪亮的科研创新点,更坚信它将造福人类的伟大使命。而要想这粒小小的药片走出实验室,实现它的价值与使命,王杰第一次意识到要做好创业的准备。
■本报通讯员许晓凤记者温才妃
小药片“美她司酮”,静静地躺在福州大学肿瘤转移预警和预防中心实验室的小瓶子里,它以创意组第二名的成绩捧得第三届中国“互联网+”大学生创新创业大赛的金奖。作为福州大学肿瘤转移预防研究的重大成果,仅2016年,该中心实验室在肿瘤转移的药物干预领域发表的论文总数就占全球总数的36%。
完美的实验数据,顶尖的课题论文,作为高校的科研成果,小药片“美她司酮”的上升路几乎可以告一段落。但它的发明人、“千人计划”入选者、教授贾力和博士生王杰,以及项目组其他的小伙伴们却有些不甘心,不忍它被束之高阁。只因这粒看似普通的小药片是眼下癌症预防转移的重大新药,未来每年至少可以挽救300多万名癌症转移复发患者。
一颗希望的种子
人们常常“谈癌色变”,然而原发性肿瘤本身并不可怕,今天的外科微手术和放射治疗可以精准地切除未转移的局部肿瘤,可怕的是肿瘤手术后的再转移。而预防高风险期的肿瘤转移,一直是全球性肿瘤治疗理论技术和产品的空白。
福州大学开辟了基于血中循环肿瘤细胞及其代谢物的肿瘤转移药物预防领域。这一特色领域旨在证明“肿瘤的发生是不可预防的,但肿瘤的转移是可以预防的。肿瘤转移后是极难治愈的,但肿瘤转移前是可以预防的”。而“美她司酮”便是这一理念的重要研究成果。
值得注意的是,小药片“美她司酮”是基于传统避孕药米非司酮加以优化而来,属于具有自主知识产权的“老药新用”。大量的动物实验证明,这粒小小的药片在结肠癌、肺癌、乳腺癌、子宫癌等10余种肿瘤模型中,抑制肿瘤转移效果明显,综合抑制率高达95%以上。而与传统抗癌药物相比,它研发周期短,药物的成药性、安全性、可预见性强,大大地降低了研发和投资风险。
2015年“美她司酮”完成药物临床前的研究工作,顺利入选国家重大新药创制备选库,成为国家Ⅰ类创新药。
而要走出实验室,走向市场,造福患者,小药片“美她司酮”至少还需要六年的时间,需要前前后后8000多万元的投入,面对着六年临床试验可能的变数,与对于高校科研工作者而言的巨额投资,作为小药片的发明人,贾力和王杰没有丝毫退缩。在这群全年无休,穿着白大褂只知埋头实验的师生看来,“相比于这粒小小药片未来可能的巨大社会贡献,这些变数与投资都不算什么”。
使命感让这群人有了执着而单纯的想法:绝不能让“美她司酮”这粒小小的药片停止前进的脚步。
创业者王杰:不能就这么停了
王杰的第一次谈判是2015年末。当时还是硕士生的他,匆忙地从实验室出来,脱下白大褂,穿着一身学生休闲服,就坐在了第一批投资者面前,像面对项目组内的导师、同学一样,他侃侃而谈地和对方介绍着他的课题与事业——“美她司酮”的原理、前景,谈到未来对于人类的贡献,这个20岁出头的年轻人,竟然有些亢奋和激动。
“如何保证投资人的收益?作为生物医药未来临床实验的变数如何把握?如果失败了,我们的投资怎么办?”几个简单的问题,把满腔热血的王杰给问懵了。投资无望,意味着小药片“美她司酮”完成实验室使命之后就要被束之高阁,和众多其他的高校科研创新一样,成为实验数据、论文资料后被放置一边。
“不能让它就这么停了,太可惜了!”作为临床医药出身的王杰深知“美她司酮”身上闪亮的科研创新点,更坚信它将造福人类的伟大使命。而要想这粒小小的药片走出实验室,实现它的价值与使命,王杰第一次意识到自己不仅要成为一名优秀的科研工作者,更要做好创业的准备。
热爱让这名年轻人开始奔走在实验室与投资谈判的路上,这是一个“痛并快乐”的过程,痛来自一次次的碰壁,而快乐来自自己对“美她司酮”未来的坚信。从2015年底,为了让“美她司酮”找到后期临床试验的投资,王杰带着项目团队先后与二十几家企业或投资人洽谈,而参加创新创业大赛,对于项目团队的师生们来说,收获的除了奖牌,最重要的是有更多的人知晓“美她司酮”,了解“美她司酮”。而知晓与了解,对于迫切要走出实验室的“美她司酮”来说,是踏出去的第一步。
一年多的奔波,虽然辛苦,但总算有所进展,目前项目团队已经与浙江某制药公司取得中试生产的合作意向,眼下正与福州某三甲医院就药物未来的临床实验工作作深入探讨。
导师贾力:效益不仅是金钱上的
如果说在王杰看来,小药片“美她司酮”是他的课题与事业,那么对于从事抗癌药物研究三十多年的贾力来说,“美她司酮”则是他一生的心血和价值体现,而他则是“美她司酮”前进路上保驾护航的引路者。
作为诺贝尔生理学或医学奖得主、著名药理学教授罗伯特·弗奇戈特的得意门生、美国国立卫生研究院国家癌症研究所的前高级项目主管,贾力一直致力于抗癌新药的研发和申报工作,在美国先后参与12项新药申报,领导或参与过35项新药开发项目。随着新药的投放使用,困惑也在这位科学家心里:为什么随着科研水平的提升和药物研发,癌症致死率依然居高不下?
带着困惑和对生物医药研究的执着,2011年底,贾力在福州大学创立了全球独具特色的“肿瘤转移的预警和预防中心”,而这也就是小药片“美她司酮”的家。中心负责研发各种能可靠预警和有效干预原发性肿瘤在手术后再转移的技术和产品,从而解除人类对肿瘤转移的恐惧,大规模地降低人类因肿瘤转移造成的死亡。
而要从实验室束之高阁的瓶子中走出来,面对着未来临床试验的变数和巨额的投资,“如何保证投资人的收益”,对贾力这位一生奉献给生物医药创新的科学家来说,回答简单而坚定,“效益从来不仅是金钱上的,‘美她司酮’未来的社会效益足以让它获得有情怀、有责任的投资者青睐”。
这就是小药片“美她司酮”的故事,未来向前的每一步,需要创业者王杰的东奔西走,也需要贾力在科研创新上的引领,更需要社会与企业的相助相伴。
《中国科学报》(2017-10-17第8版科创)
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