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

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

What is the best method to evaluate and measure non-circulating estrogen levels in mice ?

Could anyone suggest me the best procedure to measure the circulating estrogen levels in mice ? I came across the general methods where it can be measured in serum or urine but a detail suggestion...

22 February 2021 4,352 2 View

Hi..how to isolate crude HDL (all subfractions) from human serum by size exclusion chromatography?

I would like to know about isolation of crude HDL from serum by SEC. Is it possible to perform with a serum sample volume of

19 February 2021 5,576 2 View

Which is the best wavelength to study aerosol optical depth(AOD) related to climate reseach,which criteria must be adopted in choosing the wavelengh?

We are doing a study on impact of aerosol on precipitation. Which satellite data and which wavelength will be the best. or can some one suggest a criteria to be adopted to choose an instrument...

12 February 2021 6,324 3 View

How do we calculate 95% confidence intervals for RMSE?

I have two datasets (measured and modelled). I want to calculate 95% confidence intervals for RMSE. Cany anyone please help me with this? Thank you in advance

04 February 2021 5,439 6 View

Queries regarding the rain flow counting algorithm?

02 February 2021 9,790 2 View

Helicoverpa armigera eggs are not hatching in lab condition, why?

Dear RG friends, I am providing a complete H. armigera diet for larvae and maintaining the eggs in the growth chamber as well as in-room conditions. But since three months, eggs laid from the...

21 January 2021 5,546 32 View

Doubt in DFT geometry preparation?

Hi all, I am trying to perform some ab inito DFT calculations using cp2k code. I have a unit cell of 120 atoms and would like to know how to refine or reduce the number of atoms in the unit cell...

20 January 2021 6,024 3 View

Why am I getting 2 bands for Gibson Cloning Vector PCR?

Hi, I designed Gibson primers (2 primers for fragment and 2 primers for vector) using snapgene. Then I ran PCR separately for Fragment and Vector. Fragment DNA was found to be the correct sized...

11 January 2021 9,500 2 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Does anyone know, can i do cryosectioning of whole aorta after oil red O staining and use the cryosection parts for further analysis?

I would like to quantify the lesions on aorta. I want to take image of atherosclerotic plaques formation in aorta by using O red Oil Solution. After the staining, what will be the storage...

16 February 2021 5,876 2 View

What is the proper in vitro culturing method to get information about the activation of innate immune cells by VLPs?

Dear All, I am interested in studying the response of immune cells against virus-like particles (VLPs). I used the PBMC in vitro assay and treat it with the VLP using PHA as a positive ctrl while...

16 February 2021 1,337 2 View

Why does flow deformation occur at the test line and control line on nitrocellulose membranes (LFIA)?

I use 1% BSA in 10 mM phosphate buffer (pH 7.4) solution when striping my nitrocellulose membranes with antigens and antibodies. However, I always observe deformed flow when my sample or running...

16 February 2021 4,224 3 View

Can the viability assays (ex: MTT/CCK-8) measure cell proliferation? or are they solely viability assays?

I want to conduct a proliferation assay to test a chemotherapy combination on a cancer cell line. At first, I thought about the MTT assay. But now, all sources are saying that these do not measure...

14 February 2021 3,142 7 View

GST assay in microplate?

I'd like to know if anyone can provide me with a protocol for traditional GST enzymatic assay (with GSH and CDNB as substrates) in a 96 well microplate, for cell line and/or C.elegans...

11 February 2021 1,328 3 View

What lysis buffer is compatible with Bradford assay?

Hello! I am planning to use Bradford assay to measure protein content in PBMCs lysates. The problem is that Bradford assay doesn't seem to be compatible with Tris-based buffers or SDS. Are there...

10 February 2021 1,797 5 View

For PC-3 cell line. How long before irradiation do I have to synchronize my cells to G0/G1?

I am trying to do a clonogenic assay for PC-3 after irradiation. I have to synchronize my cells by serum starvation but there's a optimal time I have to do that before irradiation. been looking in...

09 February 2021 9,480 1 View