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 R2061, R2063) 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), ≥6 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: 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.
Q8: 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.
Q9: Is it possible to isolate DNA with the Direct-zol RNA kits?
Direct-zol DNA/RNA (D2080) kits can isolate DNA from TRIzol
Q10: 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.
Q11: 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.
Q12: 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.
Q13: 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.
“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 | |
|---|---|---|---|---|
| E1010-1-16 | DNA Digestion Buffer | 16 mL | $29.00 | |
| R2050-1-50 | TRI Reagent | 50 ml | $70.00 | |
| R2050-1-200 | TRI Reagent | 200 ml | $219.00 | |
| E1010-1-4 | DNA Digestion Buffer | 4 mL | $15.00 | |
| W1001-10 | DNase/RNase-Free Water | 10 ml | $19.00 | |
| W1001-4 | DNase/RNase-Free Water | 4 ml | $12.00 | |
| R2050-2-160 | Direct-zol RNA PreWash (Concentrate) | 160 ml | $166.00 | |
| R2050-2-40 | Direct-zol RNA PreWash (Concentrate) | 40 ml | $42.00 | |
| C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
| R2062 | Direct-zol RNA Microprep | 200 preps | $545.00 | |
| R2060 | Direct-zol RNA Microprep | 50 preps | $175.00 | |
| C1004-50 | Zymo-Spin IC Columns | 50 Pack | $53.00 | |
| R1003-3-48 | RNA Wash Buffer | 48 ml | $105.00 | |
| R1003-3-12 | RNA Wash Buffer | 12 ml | $30.00 | |
| E1010 | DNase I Set | 250 U | $56.00 |
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
就好像撒一撮芝麻在培养基那么大的地方上，我们不能知道它会怎么落。
原代细胞产品专家齐氏生物提醒，原代细胞培养注意事项，供参考：
1、实验材料要新鲜，从活体分离材料后要低温保存，并尽快进行细胞分离实验。
2、无菌操作。操作时用的培养液，可加平时细胞培养液5倍含量的青链霉素。
3、用酶法分离细胞时，注意酶液的浓度和控制消化时间。
贴块法分离细胞时，注意动作要轻柔，不要伤到细胞组织，组织块边缘尽量平整有利于细胞游离。
4、培养液的选择。不同的细胞有对培养液中营养的要求不同，根据所分离细胞的特性选择。
不知道楼主明白没有,意思是说培养基上的癌细胞可能过密,导致营养缺乏,无法满足生长分裂的需要,所以进入G0期,暂时不生长分裂.
3,MHC基因位于第六号染色体上,MHC存在于人类和哺乳动物的细胞表面.根据MHC所编码的分子结构和功能不同而将其分为1,2,3类?.1,2类分子结构相似,主要功能是向T细胞呈递抗原,是被T细胞识别的重要靶分子.1类存在于除红细胞以外的机体所以细胞膜上,2类分子仅存在于成熟B细胞,一些T细胞和抗原呈递细胞膜上,3类分子不具备上述功能,属于血清蛋白.
现在楼主应该明白了,MHC是不参与抗原抗体特异性结合的,是专门用来呈递抗原的.
细胞株(cell strain) 是通过选择法或克隆形成法从原代培养细胞中获得具有特殊性质或标志物的细胞称为细胞株。一般认为，细胞株是用单细胞分离培养或通过筛选的方法，由单细胞增殖形成的细胞群。细胞株的特殊性质或标志必须在整个培养期间始终存在。
细胞系(cell line) 是原代细胞经首次传代成功后即为细胞系。泛指一般可能传代的细胞。其中能够连续传代的细胞叫做连续细胞系或无限细胞系，不能连续培养的称为有限细胞系。大多数二倍体细胞为有限细胞系。由原先存在于原代培养物中的细胞世系所组成。如果不能继续传代，或传代次数有限， 可称为有限细胞系(finite cell line)， 如可以连续培养， 则称为连续细胞系(continuous cell line)， 培养50代以上并无限培养下去。人类肿瘤细胞，在体外培养半年以上，生长稳定，并连续传代的即可称为连续性株或系。
不知道楼主明白没有,意思是说培养基上的癌细胞可能过密,导致营养缺乏,无法满足生长分裂的需要,所以进入G0期,暂时不生长分裂.
3,MHC基因位于第六号染色体上,MHC存在于人类和哺乳动物的细胞表面.根据MHC所编码的分子结构和功能不同而将其分为1,2,3类?.1,2类分子结构相似,主要功能是向T细胞呈递抗原,是被T细胞识别的重要靶分子.1类存在于除红细胞以外的机体所以细胞膜上,2类分子仅存在于成熟B细胞,一些T细胞和抗原呈递细胞膜上,3类分子不具备上述功能,属于血清蛋白.
现在楼主应该明白了,MHC是不参与抗原抗体特异性结合的,是专门用来呈递抗原的.
哪种要好，有什么要求吗？接种多少代的细胞？
1.培养基最好是培养原代细胞专门的培养基或说是配套的培养基; 2.关键还是生长因子等的浓度; 3.如果需要的话,需要明胶包被.
2.关键还是生长因子等的浓度；
3.如果需要的话,需要明胶包被.
