Please suggest me which universal primers should I use to identify fungi upto species level?

10 November 2016 8,952 4 View

Can we amplify gene of interest from cDNA or mRNA using single degenerate primer?

I want to clone my gene of interest from a eukaryote.....I have a single 5' degenerate primer available to me. can I use single 5' degenerate primer from my amplification and oligo T as my reverse...

01 February 2016 5,835 4 View

Can anyone suggest as to why I am not able to see the expression of 45KDa protein in pET22b?

I have cloned my genes of interest in pET22b and still not able to see the expression in BL21(De3)plys.

10 November 2014 7,866 8 View

Hello everyone....

I am looking for a post doctoral position in Microbiology and currently i am pursuing post doctoral fellow in Department of Microbiology University of KwaZulu Natal (Westville campus) Durban South...

06 July 2014 1,800 1 View

Can anyone help with phosphate estimation problems?

I am getting the dark blue colour even before adding phosphate to my reagents when estimating phosphate with ascorbate method. Could someone troubleshoot this for me?

04 May 2014 3,274 5 View

Does anyone have any suggestions for cloning the entire genome of an RNA virus?

I want to clone the whole viral genome from a water sample from which I have concentrated the virus and done DNA and RNA isolation. I want to find the diversity of viruses and therefore would like...

04 May 2014 6,611 2 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

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 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

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View