How to deal with the recudtion of amplification efficiency of indexed primers?

Hi all, I am now working with ITS2 sequencing library preparation. I had observed that after introducing adapters, linkers and barcodes to a primer, the indexed primers have a much lower...

27 January 2021 5,045 3 View

Are there any DNA intercalators that do not have fluorescence?

I understand several normal DNA intercalators, such as ethidium bromide, acridine, SYBR et al. would emit fluorescence. Is there any kind of DNA intercalator that do not have fluorescence?

25 December 2020 688 3 View

How to interpret the centrality indices in network analysis?

I learned that in order to interpret the centrality indices, the CS coefficients resulted from a bootnet test are preferably above 0.5 and above 0.25 is acceptable. In my network, the CS...

29 October 2020 3,545 3 View

How to create a graph of psychopathology symptoms when conducting network analysis in R?

I've been working on the network analysis of adolescent depressive symptoms. I've estimated the network, the centrality indices and network stability using packages qgraph and bootnet. Then I want...

22 October 2020 5,591 3 View

How to interpret the results of NetworkComparison Test when conducting a network analysis in R?

25 September 2020 9,936 2 View

Does anybody know how to use the Lagranto model?

These days I wanted to run the Lagranto model to track water vapor transport path. But I got some trouble in installing an available version of Lagranto model. I attempted to use "conda install -c...

04 July 2020 5,417 4 View

Abaqus & Fortran complier linking problem ?

Hi, I am trying to run a vumat subroutine, but there is some abaqus-fortran linking issue. I use abaqus 6146 Micrasoft visual studio 14.0 parallel studio XE 2016 I added these command in my .bat...

25 February 2020 5,698 2 View

Health informatics: Relationships between industry development and academic research?

In digital health domain, is the academic research that leads the development of the industry OR is the industry development that impacts the associated academic research ?

10 June 2019 5,214 6 View

Fluorescence Polarization Assays?

When I titrated fluorescence probe by Protein A alone or by Protein B alone, it do not should any binding signals, mP do not change. However, when I titrated the same fluorescence probe by...

18 March 2019 2,468 3 View

How does esiRNA from Sigma compare to Dharmacon's SMARTpool siRNA?

I would like to know information about esiRNA form Sigma for the following points: 1. transfection protocol; 2. knockdown efficiency; 3. off target effects; 4. toxicity. Anyone would like to share...

31 May 2018 9,661 2 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

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

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Errors in DNA sequences?

I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...

24 February 2021 4,397 3 View

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View