Anyone having experience with Genecopoeia All-in-One™ qPCR Primers for mouse transcripts?

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What is appropriate collagen induced arthritis (CIA) immunization protocol for DBA1/J mice?

I am planning to run a kinetic CIA experiment using male DBA1/J mice and I would like to see change in expression of gene of interest at various stages of disease. I have been reading about the...

03 April 2015 3,386 4 View

How to avoid saturation of positive control protein bands in Western blotting?

I am wondering how one can avoid saturation of loading controls while detecting low abundance protein of interest. I usually load 15-20microgram/lane of nuclear fraction from tissue homogenate to...

10 November 2014 1,073 5 View

Is it possible to have a candidate gene with no coding SNPs but differential levels of methylation?

I am working on a genetic region (quantitative trait locus) that was identified in linkage analysis between two mouse strains that differed in response to induced arthritis. I am trying to...

09 October 2014 8,536 9 View

Is CD3/CD28 stimulation and ConA stimulation activating T cells via different mechanisms?

I am trying to compare different groups of mice for T cell proliferation responses. I use unsorted splenocytes and stimulate them with different concentrations of antiCD3/CD28 antibodies and ConA....

09 October 2014 8,194 12 View

Could someone help with an in-vitro osteoclastogenesis assay protocol (from whole bone marrow culture)?

I'm planning to set-up an in-vitro osteoclastogenesis assay from mouse bone marrow culture, followed by TRAP staining. Could someone share a detailed, working protocol for that? Any suggestions...

06 July 2014 6,568 16 View

Any suggestions for a choice of growth and differentiation medium for bone-marrow stromal cells?

I am going to grow mouse BMSCs in-vitro and I want to differentiate them into osteoclasts. I am very new to this so I have been looking around for various protocols and publications for this kind...

03 April 2014 4,294 4 View

What is the importance of intronic SNPs in terms of structure/function of protein?

I am working with mouse models of RA and we have recently sequenced the whole genome of two strains to reveal genetic differences. Now I have a long list of SNPs for the 2Mbp region (relevant for...

02 March 2014 7,591 21 View

Nanodrop or BCA/Bradford assay?

I am interested to know if it should be okay to quantify proteins in a lysate by using nanodrop for purpose of Western Blotting. I have seen most of the protocols with either BCA or Bradford...

11 December 2013 1,803 7 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Research Methodology - Impact of Corporate Reputation on Stakeholders Behaviors?

Please can anyone support with the survey questions based on RQ measures and propose how to do it in FMCG industry and include as well the role of brand equity Thanks

06 August 2024 949 0 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Please explain how the plastic input value should be considered from the true stress-strain curve for the bilinear elastoplastic material model ?

I am working on Abaqus/Explicit(Quasistatic ) for the deformation of the auxetic structure model. Please explain how the plastic input value should be considered from the true stress-strain curve...

05 August 2024 454 3 View

"A Markov-like Model for Patient Progression"?

A Markov-like Model for Patient Progression" Markov Chain Monte Carlo (MCMC) Markov Chain Monte Carlo (MCMC) is a powerful computational technique used to draw samples from a probability...

05 August 2024 10,079 0 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

How to report results of Generalised Linear Mixed Models in a journal article?

Hi everyone, If you have written or come across any papers where Generalised Linear Mixed Models are used to examine intervention (e.g., in mental health) efficacy, could you please share the...

04 August 2024 4,130 4 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View