What to do if the gene we are interested does not give positive staining?

Hi , I want to know that if I hypothesize that Gene A promoting chemoresistance in PDAC but what to do if I do not get any positive staining in my sample? What measure will I have to take which...

24 September 2020 7,370 3 View

If I have to select between Gasteiger and AM1-BCC charge types, what is the best option here for my ligand?

If I have only two options in front of me to select either Gasteiger or the AM1-BCC, so based on which parameters or rules I can select the most appropriate charge scheme during minimization step...

18 July 2020 8,763 1 View

What impact would be there on RNA-SEQ experiments if we do not use any RNA Spike-In controls (like ERCC or SIRVs)?

Hi everyone, if I do not use spike-in RNA isoform controls for a customized RNA sequencing experiment (not the whole transcriptome, just based on few specific genes we are interesting), so is...

08 February 2020 7,190 2 View

What is the minimum and maximum cDNA input from FFPE samples in library preparation phase to perform the RNA SEQ by Ion Proton?

Dear All, 1 ug of permissible quality FFPE RNA is transcribed to cDNA through SSIV VILO RT kit. I have Qubit v.4 device, which can measure cDNA to a good extent. The cDNA ranges from 25-100 ng /...

11 November 2019 2,009 1 View

What are the conditions of different Regression Models?

i want to ask about the appropriate model for the prediction of 4 type of learning styles with the help of background variables of the respondents In my case the dependent as well as the predictor...

19 December 2018 7,237 3 View

Is there any undesirable effect of bacterial RNA load of normal intestinal flora present on human colon samples for the gene expression analyses?

It is a known fact now that interactions between bacterial (normal flora) and colon cancer cells influence the transcription profile of host cells or enterocytes. So, how can I avoid or minimize...

11 October 2018 9,483 1 View

For customized mRNA sequencing, can I omit the rRNA removal procedure step, which is offered by several commercial kits?

If I am interested in a customized mRNA-only Sequencing (RNA-Seq) based on few transcripts (not the whole transcriptome), and after having satisfied & permissible RIN values for my extracted...

21 September 2018 1,804 3 View

Does "No Primer Dimer Formation" guarantee the absolute utilization of entire primer content (F & R both) in a PCR reaction?

Dear all, in a PCR (conventional & realtime both), if we achieve our desired product (GOI) and along with, there is also no primer dimer (PD) formation seen on the gel and melt curve, does...

02 November 2017 9,949 3 View

Why does the HRMA (High Resolution Melt Analysis) cannot detect homozygous variants properly than heterozygous ones?

Any explanative thoughts or information? Will the 2nd or 3rd melt of products attribute to some improvement for their detection?

20 October 2017 8,562 3 View

Is that logical? Two strains of the same bacteria with too much different growth kinetics?

Greetings! I'm working on two strains (PAO1 and PAK) of the same bacteria, Pseudomonas aeruginosa. I have obtained the growth curves of the two strains separately to have the OD vs CFU/ml curves...

02 May 2017 8,123 2 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 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

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View