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 |
| Sample Inactivation | TRI Reagent® (provided with R2051, R2053) 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 | 50 µg RNA (binding capacity), ≥25 µl (elution volume) |
Q1: Is DNase I available for individual purchase?
All kit components are available for purchase separately.
Q2: 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.
Q3: 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.
Q4: 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.)
Q5: 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.
Q6: Is the DNase-I treatment necessary?
If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.
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.
“No phase separation was needed, but you still had the benefits of a Trizol extraction. No need to precipitate and resuspend samples, which means less sample loss during purification.”
-Adina B. (University of Guelph)
“This kit is amazing, I"ve got a gel comparing the lack of gDNA as shown in the advertising pamphlet. What can I say, except: I love this product!“
-R.K. CSU
“Direct-zol is the most excellent kit for RNA isolation that I ever used in the past 20 years.”
-H.Z. (Harvard Medical School)
Read More| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| E1010-1-4 | DNA Digestion Buffer | 4 mL | $15.00 | |
| E1010-1-16 | DNA Digestion Buffer | 16 mL | $29.00 | |
| R2050-1-200 | TRI Reagent | 200 ml | $219.00 | |
| R2050-1-50 | TRI Reagent | 50 ml | $70.00 | |
| C1078-50 | Zymo-Spin IICR Columns | 50 Pack | $55.00 | |
| R1003-3-12 | RNA Wash Buffer | 12 ml | $30.00 | |
| R1003-3-48 | RNA Wash Buffer | 48 ml | $105.00 | |
| E1010 | DNase I Set | 250 U | $56.00 | |
| R2050-2-40 | Direct-zol RNA PreWash (Concentrate) | 40 ml | $42.00 | |
| R2050-2-160 | Direct-zol RNA PreWash (Concentrate) | 160 ml | $166.00 | |
| C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
| R2050 | Direct-zol RNA Miniprep | 50 preps | $180.00 | |
| R2052 | Direct-zol RNA Miniprep | 200 preps | $555.00 | |
| W1001-6 | DNase/RNase-Free Water | 6 ml | $15.00 | |
| W1001-30 | DNase/RNase-Free Water | 30 ml | $22.00 |
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引物是一小段单链DNA或RNA，引物可以做为DNA复制开始时DNA聚合酶的结合位点，在细胞外的条件下，只有通过引物，DNA才可以开始进行复制。
引物是人工合成的两段寡核苷酸序列，一个引物与感兴趣区域一端的一条DNA模板链互补，另一个引物与感兴趣区域另一端的另一条DNA模板链互补。
启动子和引物两者的区别：
启动子是DNA分子上能与RNA聚合酶结合并形成转录起始复合体的区域，在许多情况下，还包括促进这一过程的调节蛋白的结合位点，决定RNA聚合酶转录起始位点的DNA序列。引物是一小段单链DNA或RNA，引物可以做为DNA复制开始时DNA聚合酶的结合位点，在细胞外的条件下，只有通过引物，DNA才可以开始进行复制。引物是人工合成的两段寡核苷酸序列，一个引物与感兴趣区域一端的一条DNA模板链互补，另一个引物与感兴趣区域另一端的另一条DNA模板链互补。
启动子是位于结构基因前的非编码区对转录起调控作用的一段DNA序列。引物则是游离于DNA复制的模板链之外的对DNA复制起调控作用的一小段单链DNA或RNA。
在这里我向大家隆重推荐一个在线的引物设计软件，是斯坦福大学的，我认为是最好的在线引物设计软件，我用它设计接近100多对引物，可以说是屡试不爽。它用起来也很简单，最重要的是效果非常好，考虑的因素也非常的多。
先不说那么多了：网址：http://www.yeastgenome.org/cgi-bin/web-primer
下面我继续图示。
帖子发表之后很多战友PM我，让我帮设计引物，其实这很让我失望的，我的本意是让大家掌握这种方法，而不是告诉大家我是一个专家，其实我水平也有限，所以我不会帮大家设计引物，我认为这是科研工作的一部分、基本功，祝大家好运、实验顺利。
帮忙设计这段DNA序列的2个引物，forwardprimer还有reverseprimer，要只含有6个bp
麻烦写详细3端和5端，可以的话写下设计方法，感激不尽啊
刚开始学很是困惑，老师也没详细讲。。。
新手一枚，看到论坛里也有许多人问过，但没找到比较确切的回答，还是不是很理解，做芯片给出的lncRNA的名字是XLOC_009167，这个名字在ncbi里面都找不出来，我该怎么样去获得这个rna的信息呢？
需要在片段5‘端和3’端分别设计引物去扩增。
所以，对应的上游5‘端，我们习惯叫做”上游引物“，即forward primer，简称for；
对应序列的3’端，我们叫做下游引物，reverse primer，简称rev。
非要说差别，可能就在测序引物是让片段线性增加，PCR引物是让片段指数增加。
请教各位高手：
1.本人最近用权威文献中的引物PCR，使用前在NCBI上BLAST了一下，可是并没有结果。这个引物可用吗？
2.自己用OLIGO7设计了一对引物，评分良好，BLAST特异性好。可是实际一直P出某条非目的条带。退火温度也按TM值调节过了。实在想不出什么原因。
3.使用NCBI上的PRIMER-BLAST设计的引物，在NCBI自己的BLAST却找不到目的片段，这是为啥？引物可用吗？
谢谢！
