Does anyone know of a more affordable alternative to Mito-Tracker, that is well retained in fixed cells?

I have in the past worked with the MitoTracker dies. They are rather expensive and that is the bit I am hoping to avoid. I would really appreciate it if anybody can suggest a mitochondrial...

05 November 2020 5,701 1 View

What is a good ligation strategy with compatible ends EcoRI and MfeI in the vector DNA and the insert?

I used to do this kind of thing and now have forgotten strategy please help and be specific. The vector is now linear having been digested with EcoRI and MfeI, then treated with CIAP, to prevent...

23 May 2020 2,377 1 View

Could anyone send me a FileMaker template to create a biological collection (bacterial strains, mutants, etc)?

I am planning to create biological databases for yeast, bacterial, strains and plasmids. I want to use FileMaker pro but I am a new user/ A template would be of great help to see how I can modify...

20 October 2018 6,708 1 View

Can I set up a plate for qPCR using Power SYBR Green PCR Master Mix #4367659 and run it a day later?

Catalog number: 4367659 Power SYBR Green PCR Master Mix. This is NOT POWER UP that I am asking about. Can I set up a plate Monday the 28th in the afternoon and run it Tuesday the 28th in...

24 May 2018 8,588 5 View

Does anyone have a tried and true protocol for transfecting RAW cells with plasmid DNA for protein expression?

02 June 2016 7,457 0 View

Does anyone have a complete protocol for generating total cell lysate for proteomic analysis from adherent tissue cultured cells?

04 May 2016 3,844 2 View

How can I image live cells in 3D systems and subsequently maintain them in culture?

I am growing bladder cancer cells in 3D models in vitro and I wish to visualize them using some form of real-time imaging which will still allow me to grow them and use them for protein/RNA...

11 May 2014 853 7 View

*CRY file for Molybdenum

Does anyone have a *CRY file for molybdenum or know where I can get one from the web, please. I am trying to EBSD a moly, titanium, iron alloy.

01 January 1970 5,127 3 View

Is there a protocol to establish a stably transfected macrophage cell line, preferably in mouse?

01 January 1970 5,394 1 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

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

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

How does the injection volume impact LLOQ and ULOQ in regression analysis (LC-MSMS)?

Hello, I have run into an issue that has sparkled a debate at work: is the LLOQ/ULOQ impacted by injection volume under the following circumstances: 1) Same dilution factor of all samples 2)...

23 February 2021 9,159 3 View

RNA degraded after good nanodrop readings?

23 February 2021 8,850 2 View