23 Questions 13 Answers 0 Followers
Questions related from Iddo Magen
Hi, I have expression levels of 92 miRNAs (after filtering out low-abundance miRNAs) in 6 patients X 5 time points during drug treatment (baseline, day 7, day 14, day 21, day 30). Now, I want...
14 March 2024 6,433 0 View
Hello, I have 5 blood samples each from 6 patients treated with a drug: on day 1 (before treatment), 7, 14, 21 and 30. In these samples I have measurement of both the drug concentrations and...
07 March 2024 6,624 2 View
I have Trizol LS that was manufactured in 2017 and was stored at RT.
16 May 2023 8,140 3 View
Hello I accidently forgot to remove traces of 70% ethanol used to wash beads, and did the elution for my libraries with water right away. So now my libraries contain a bit of ethanol, I do not...
13 January 2022 4,890 1 View
Hello, Does anyone know about cheap services for library preparation of small RNAs from biofluids? It may also include RNA extraction/sequencing and can be outside North America/Europe. Thanks
31 August 2021 6,501 3 View
Hi I am going to slice paraffin embedded spinal cord. I have two questions: 1. How do I put the SC in the paraffin block so that it will be in coronal orientation? It is not trivial as the part...
14 August 2020 7,822 2 View
I am going to do in situ hybridization for adult mouse brain and spinal cord. I will post-fox the tissues in 4% PFA ON and then embed in paraffin. Should I also perfuse the mice before with PFA? I...
11 August 2020 4,233 6 View
Hi I wish to perform ddPCR for miRNAs in blood plasma. I have a kit for preparing cDNA with a universal RT primer that uses poly-A tail as a template, and a Taqman probe for qPCR that is called...
14 June 2020 3,677 3 View
Hello, I need to collect blood plasma from animals for RNA extraction, and I want to add Qiazol and freeze quickly in dry ice to preserve the RNA. The problem is that in the procedure room where...
27 April 2020 8,349 0 View
Hi , i performed multivariate Cox regression analysis and one of the covariates had a p-value of 0.08. since it is a replication study and i have a hypothesis, I can have a directional...
03 May 2019 7,552 2 View
Hi, I wish to validate NGS data of miRNA sequencing from human plasma samples. The obvious way to do it is of course Taqman, but there are two different assays: the 'traditional' one in which...
16 January 2019 540 0 View
Hi everyone, I did RNA-seq for small RNA, and now I want to confirm the results with qPCR. What is the best way to find reference miRNA genes from the NGS data, for the normalization of the...
13 January 2019 1,748 3 View
Hi, i performed NGS for small RNA and ran DESeq on the data. there is one miRNA that is upregulated by 2-fold in the patients compared to healthy controls. However, when i look at the raw...
05 July 2018 6,344 3 View
Hi, I am performing a biomarker study and I have a certain AUC value after performing machine learning algorithm (radom forest). Is there a way to tell how many additional samples (from control...
17 June 2018 5,783 3 View
Hi, I need to isolate plasma from blood for RNA extraction afterwards. Does it have to be in a cold centrifuge? Do you need in general to isolate plasma in a refirigerated centrifuge or is it...
27 May 2018 1,459 4 View
Hi, I have mice with a gene deletion in the brain, because the Cre deleter is expressed under the Nestin promoter. On what embryonic day can I expect Nestin to be expressed ? I suspect that the...
17 October 2017 3,347 1 View
Hi, Does anyone know if genotyping mouse embryos follows the same protocol as tail genotyping? i.e., I have to take one piece of the embryo and incubate with lysis buffer + proteinase K and then...
17 October 2017 5,381 0 View
Hi, I got these results in tape station (attached). The small RNA library size should be around 173bp, so I think that libraries 28, 31, 34, 36, 37 also have an adapter dimer. I am going to...
02 October 2017 620 1 View
Hi, I wonder if a single stranded DNA can be polyadenylated or only single stranded rna?
19 June 2017 8,556 1 View
Hi, I would like to extract RNA from biofluids, and we have a home-made protocol that I got from a collaborator, that uses Zymo spin columns. However, we have bunches of miRNeasy micro minelute...
14 June 2017 8,493 1 View
Hello, If I use a PCR plate for library prep and I need the products after each step in the protocol (in my case it's polyadenylation, RT and PCR), is it safe to open the plastic foil that covers...
26 April 2017 8,641 3 View
Hello, Regarding reliability of RNA-seq data - I sequenced a serum sample with the NEB kit and a new kit from Clontech that is PCR-based and not ligation-based so it is supposed to introduce less...
22 April 2017 4,170 5 View
Hi, I have mice expressing Cre under the Nestin promoter and the floxed gene. where should I expect deletion of the gene? In the whole brain? only neurons or also glia?
11 March 2017 6,004 3 View
