Contaminated taqman probe?

More Martin Forde's questions See All

Can I use blocking peptide as positive control in Western blot?

I'm just curious if blocking peptides can be used in this fashion? Does the conjugation of the blocking peptide with the primary antibody stop the immunoreactivity with say a flouro-tagged...

31 December 2015 5,676 5 View

MDA-MB-231 advice?

I'm looking to do a time course realtime-glo vitality assay on MDA-MB-231 in a 96-well plate. I plan on doing 4 different seeding concentrations to assess linearity of signal, however there is...

05 June 2015 1,888 4 View

How can I perform SiRNA concentration scaling? by nM or pmol per num of cell / surface area?

I'm looking to scale up my siRNA reverse transfections from a 6-well to 384-well plate for adherent SH-SY5Y. Should I be scaling the siRNA concentration by surface area in the well, or by the...

31 December 2014 4,874 6 View

How can I explain great siRNA transfection efficiency but no knockdown with validated positive controls?

I'm performing reverse transfection on SH-SY5Y using Viromer Green, and I get wonderful transfection efficiency (90+%). See picture (RFP labeled siRNA fluorescence overlapped on phase contrast...

10 November 2014 7,589 7 View

Can anybody recommend how much Blasticidin should be used for selection of SH-SY5Y?

We are electroporating a pcDNA6.2 plasmid contain miR RNAi constructs into SH-SY5Y. I plan on selecting these cells with blasticidin, and the literature suggests closer to the 10ug/ml mark for...

09 October 2014 6,550 3 View

Is anyone familiar with electroporation?

I thoroughly pipette my cells prior to electroporating so they should form a monolayer when plated. Though every time I electroporate, they clump. I'm not entirely sure why. When I passage, they...

08 September 2014 6,425 3 View

Shsy5y immunostaine troubleshooting

cells plated

07 August 2014 2,284 0 View

What is a good number of SH-SY5Y to freeze down for long term storage between 1-12mo?

What is a good number of SH-SY5Y to freeze down for long term storage between 1-12mo? Ideally, I'd like to initially plate in something larger like a t75 or t150, though I'm not sure if it is...

06 July 2014 4,850 2 View

What is the best method to carry out SH-SY5Y differentiation and transfection?

How do people typically go about electroporating differentiated SH-SY5Y? I was thinking that you would have to electroporate them first then recover in media supplemented with retinoic acid. Would...

06 July 2014 1,798 1 View

Can anyone share their knowledge on cell penetrating proteins?

Has anyone had any experience using CPPs to transfect small plasmids or siRNA into primary human neuron? I'm just curious to see if you would share your experiences. I'm going to be testing with...

06 July 2014 1,498 2 View

Which type of compound does lamda max of 218 indicate in a uv-vis spectrum of a partially purified compound through column and TLC?

A crude extract of fungal culture using EtOH was subjected to column and TLC and partially purified compound was obtained. UV vis spectrum of the compound/s has max absorbance at 218nm. The...

11 August 2024 9,801 2 View

Baseline drift in HPLC? What causes this?

Hello, Why do i see this baseline drift when i compare my blank (black) to the sample (blue)? Any suggestions as to why this happened? Thank you!

11 August 2024 3,770 4 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

U you think We need a website software of Blackbody radiation law expert software?

A website software of Blackbody radiation law expert software can used through the following web site. http://39.105.188.151:3000/index

07 August 2024 1,706 0 View

E.coli contamination in human RNA seq data ?

Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...

06 August 2024 807 3 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

Can a Conventional 50W visible LED Lamp Emit UV Light?

I'm researching light sources and their UV emissions. I'm curious if a 50W, 7500K conventional LED lamp emits any UV light. I found that a 200W tungsten lamp can emit up to 1.5 mW/cm² at 350 nm....

31 July 2024 3,843 7 View