Tweet
Happy new year, everyone. I am posting to receive any advice you have for me regarding an issue I'm having with RiboZero plant leaf! I am using the kit for RNA extracted from Oryza sativa. I've attached the Nano trace for DNase-treated samples and the Pico trace for RiboZero treated samples. It looks like there is basically nothing left after RiboZero, at least for the case of the first 4 samples. The last two look like incomplete subtraction, or, something else went wrong. Any tips?
-
- Gina -
This is really a question for Epicentre tech support. But, for what it is worth:
(1) We generally get extremely low yields of non-rRNA from plant total RNA compared to animal rRNA.
(2) 90% of the time when there is some protocol of this sort -- dealing with dilute solutions of RNA or DNA that produces a poor yield -- it is the result of attempting to recover that RNA or DNA via alcohol precipitation. Sadly it seems few people are taught that this this methodology is difficult and has a high failure rate in all but the hands of the most talented bench scientist. So if you see a protocol calling for you to precipitate RNA or DNA solutions at concentrations likely to be below 50 ng/ul, look for an easier method. Spend an extra few dollars on a zymo column rather than wasting weeks (or longer) trying to get the alcohol precipitation to work.
--
Phillip -
Hi Phillip, thanks for your answer. I used RNAClean Ampure beads to recover my RNA - my experiences in the past with these beads has been good so I'm not sure if this step is the problem. I am in contact with an Epicentre rep right now... will post a reply if I get some good results!Originally posted by pmiguel View Post- GinaComment
-
Hi Gina,
Maybe too late but this week I made a trial library starting the Ribozero with ~10ng! It worked except for some primer dimers, the library was there (Scriptseq V2, Epicentre) as seen by Bioanalyaer and PCR on 3 different genes. Look for my recent question (no-one replied
to see the attached curves.
Good luck,
GuyComment
-
I also succeeded in making libraries out of very little starting material.
I face another problem: The riboZero reaction is inconsistent and sometimes doesn't work at all.
These are my parameters: 2.5 µg of RNA, 8 µl of removal solution. the sample is young leaf of rice and I use the RiboZero plant leaf kit.
Anyone had difficulties?Comment
-
Kheloyar – Everything You Need to Know About Kheloyaar Login and Kheoyar Id
If you are looking for an online gaming platform that offers a user-friendly experience, Kheloyar has become a name that many users search for. Whether you're interested in creating a new account, accessing your dashboard through Kheloyaar Login, or learning how to obtain a Kheoyar Id, understanding the platform's features and account process is essential.
This guide explains everything you need to know about...Today, 01:13 AM -
Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
...07-09-2026, 11:10 AM -
Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.
There is no single reason why many patients don’t respond to treatment as expected. Cancer is...07-08-2026, 05:17 AM
Comment