I did like to know what to do to get a PCR product that will come out with a single clear band, no noise and mixed peak when the product is sequenced?

I am trying to check the polymorphisms of a particular gene from human DNA samples. A set of primer in a published publication allowed the amplification of the DNA hence permitting the analysis of...

02 March 2019 8,268 5 View

What could be the cause of noise or mix in PCR sequence result?

I subjected some genomic DNAs to PCR reaction and later sent then for sequencing to analysis for polymorphisms in my gene of interest? Some of my sequence results came out fine while some came out...

01 February 2019 9,564 7 View

Deglycosylation test: I can't understand why the bands from my deglycosylated proteins are not neat?

I carried out a deglycosylation test to see if there will be difference in the electrophoretic mobility of C57BL/6 infected-ME7 and SShpPrP Tg ( transgenic mice expressing sheep prion protein)...

01 February 2019 5,754 2 View

My transfected cells do grow at sporadic rate a week after subjecting them to antibiotic selection treatment , what could likely cause this?

I am trying to establish a stable cell line expressing a gene of my choice using a prion protein knock out cell line, Lipofectamine and Zeocin as selection antibiotics. Not ruling out the fact...

11 December 2016 8,981 7 View

What could be the likely causes of false positive or false negative results during PCR reaction?

I did tail biopsy to genotype a particular gene. The result came out positive for my desired gene but two days after using same lysed genomic DNA, it came out negative. In my confused state, l...

11 December 2016 5,573 7 View

Please l need recommendation on antibodies that can detect Chronic Wasting Diseases Prion Protein in cell lysate or brain tissue homogenate?

11 December 2016 2,281 2 View

Please l need some one to help me with protocol to establish cell line using Lipofectamine LTX plus reagent and G418 as antibiotics for selection?

07 August 2016 6,533 2 View

Please help with expert views based from experience on establishing cell lines expressing prion protein gene from any of the cervids- elk, deer ?

I need help on the most appropriate transfection approach and conditions to optimize cell transfection when establishing stable cell lines expressing any of the cervids' prion protein genes and...

06 July 2016 2,462 0 View

I did a transient transfection on Zpl neuronal cell line but the transfection wasn't successful. Please l need help on what could have caused this?

I am trying to establish a cell line using Zpl cell line- PrP knock out and LTX/PLUS lipofectamine reagent as transfection reagent.

06 July 2016 247 4 View

Can Micro RNA be extracted from tissues?

I need protocols or procedures for extracting Mi RNA from tissues in experimental animals.

04 May 2016 1,103 4 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

No title

02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 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