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

For BCA or brain homogenate preparation, what volume of Lysis buffer should be added to weighed cell pellet or tissue when lysing cell or tissue?

06 July 2016 4,526 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

What is the most Operating System (OS) which can be INSTALL on Tiny hard devices like Raspberry Pi (RPi) hardware ?

I'm targeting to deploy a mesh network and manually configure MANET routing protocols. I'm preparing scenarios, architectures, and hard devices needed to do that. Are there some step-by-step...

03 March 2021 1,931 5 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How can I integrate a routing protocol in a tiny device (e.g. Raspberry Pi)?

Hi Community, I'm facing the issue of integration/compiling a new routing protocol in a WSN simulator. The final goal is to successfully add, configure this routing protocol in hardware devices...

01 March 2021 9,332 6 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Is there any one who can provide a donation of HPV detection and genotyping kits in exchange collaboration fors?

Warm greetings My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology (on Human papillomavirus). At this stage, I have almost completed a clinical sample Cervical swabs)...

25 February 2021 1,660 3 View

Is there any one who can provide a donation of HPV detection and genotyping kits in exchange collaboration forms?

Warm greetings! My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology focusing on Human papillomavirus). At this stage, I have almost completed a clinical sample (Cervical swabs)...

25 February 2021 3,612 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View