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 3,273 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 943 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 8,409 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 8,062 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 6,442 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 4,187 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 7,470 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 1,596 7 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View

What analysis to choose with a large neste dataset and clearly skewed or kurtotic distributions?

Hello! I have a dataset of n=3000 nested within 8 countries with approximately 200 or 400 responses in each country. I originally planned to perform multilevel modelling with 4 dependent variables...

02 March 2021 6,865 1 View

How to calculate BREQ 3 questionnaire?

how to calculate BEHAVIOURAL REGULATION IN EXERCISE QUESTIONNAIRE BREQ 3 ,which contain 24 questions and each question has 5 answers (0- Strongly disagree , 1-disagree, 2- Neither agree nor...

02 March 2021 811 3 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

I am interested in medicinal chemistry field, which books are highly recommended to learn from?

I am at the beginner level in drug design and drug discovery, and I am interested to learn more about this field. Which books are highly recommended to start from and gives me all essential...

02 March 2021 8,437 2 View