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 | Microplate centrifuge, vortex |
| Sample Inactivation | TRI Reagent® (provided with R2055, R2057) 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), ≥10 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 DNase-I treatment necessary?
If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.
Q4: 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.
Q5: 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.)
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.
“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 | |
|---|---|---|---|---|
| E1010-1-16 | DNA Digestion Buffer | 16 mL | $29.00 | |
| R2050-1-200 | TRI Reagent | 200 ml | $219.00 | |
| E1010-1-4 | DNA Digestion Buffer | 4 mL | $15.00 | |
| C2003 | Elution Plate | 2 Plates | $19.00 | |
| C2002 | Collection Plate | 2 Plates | $22.00 | |
| C2007-2 | 96-Well Plate Cover Foil | 2 Foils | $10.00 | |
| C2007-4 | 96-Well Plate Cover Foil | 4 Foils | $10.00 | |
| C2004 | Zymo-Spin I-96 Plate | 2 Plates | $142.00 | |
| W1001-30 | DNase/RNase-Free Water | 30 ml | $22.00 | |
| W1001-10 | DNase/RNase-Free Water | 10 ml | $19.00 | |
| R2050-2-160 | Direct-zol RNA PreWash (Concentrate) | 160 ml | $166.00 | |
| R2054 | Direct-zol-96 RNA | 2 x 96 preps | $418.00 | |
| R2056 | Direct-zol-96 RNA | 4 x 96 preps | $675.00 | |
| R1003-3-48 | RNA Wash Buffer | 48 ml | $105.00 | |
| E1010 | DNase I Set | 250 U | $56.00 |
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1. Buffer中离子浓度过大，甘氨酸或者Tris碱有可能疏忽多加了
2.没有加相应浓度的SDS
3.电泳周围温度高，也是一个原因
加样缓冲液的主要作用是使PCR产物与其混合,使DNA沉于加样孔的底部,防止DNA跑出来.
如果你是制胶或者做电泳缓冲液的话，不用灭菌。
低离子强度时，迁移率快。但离子强度过低，缓冲液的缓冲容量小，不易维持pH恒定。高离子强度时迁移率慢，因此为了获得更快的反应速度，在缓冲容量允许的范围内，离子强度应该尽可能小。
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