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 |
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2.《Adetailedmetabolic?uxanalysisofanunderdeterminednetworkofCHOcells》
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1-s2.0-S016816561001878X-main.pdf(521.01k)
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
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。
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
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
