What is the minimum amount of sample to start an RNA sequencing experiment (for both human and mice blood samples)?

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What is the best statistical test that can deal with large data sets?

Dear All: I am wondering about the best statistical test that can deal with large data sets. I am profiling plasma microRNAs using a high throughput method and comparing the expression of those...

05 June 2016 7,590 5 View

Can anyone advise about what data to plot in the ROC curve to obtain the sensitivity and specificity of a microRNA as a disease biomarker?

Dear All: can anyone advise about what data to plot in the ROC curve to obtain the sensitivity and specificity of a microRNA as a disease biomarker? we are validating our target microRNAs...

31 December 2015 9,881 12 View

Can anyone help analysing microRNA microarray data?

I am doing an experiment to check plasma microRNA expression profile in two time points in healthy individuals by using AB openarray protocol. In some samples the raw (time0) ct values are...

04 May 2014 7,000 4 View

Using Nanodrop 2000 to detect blood sample hemolysis?

Can any one advice me on the way to perform spectrophotometric analysis on blood samples to check for hemolysis using Nanodrp 2000? We need to measure absorbance at 414 but because I am not...

10 November 2013 6,305 8 View

Normalization of microRNA profiling study in plasma samples?

I am wondering what is the best method for normalization of microRNA in RT-qPCR experiments. My samples are plasma samples so can I use a reference gene for normalization? And what is the most...

10 November 2013 5,722 3 View

Can someone suggest a sample transport in liquid nitrogen?

My work involves transportation of biological samples from the hospital to the lab. I was wondering about the best liquid nitrogen container to do that, my samples are small so a 1-2 L containers...

09 October 2013 4,300 16 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

How to compare 2 samples recruited with different sampling methods?

If I want to compare characteristics of 2 groups of patients, one group is recruited from a random sample of the general public while another group is from a sample with a fixed gender (male to...

26 February 2021 3,526 3 View

Florian Halbritter

That depends on a number of factors, most prominently sequencing platform and the precise protocol you wish to follow..

Martin Larke

I disagree with Darrell. Using the SMARTer kit form clontech we have generated libraries for sequencing from 100pg of input (10 cells in lysis buffer). This kit is very robust but takes approximately 2 days to generate libraries and is quite hands on as it requires reverse transcription and amplification of transcripts before library preparation.