How should I prepare solutions of divalent metals at neutral pH?

Hello, I would like to prepare metal solutions of 50 mM Mg, Zn, Ca, and Cu (separately) from their chloride salts, to study protein-metal-ion interaction. For this reason, I would prefer that it...

25 November 2020 1,924 11 View

What is the limit of detection for Pour Plate method?

i preformed several test using the pour plate method with no dilution, i counted 0-4 colonies per plate, inoculum 1 ml, agar-15ml, which meen 0-400cfu/100ml. my goal is to achive

26 June 2019 7,406 2 View

How to calculate the specific capacitance of graphene foil using galvanostatic charge-discharge?

The proper way is to normalize it according to the weight of the electrode in order to get the specific capacitance in F/g units? or should I normalize it according to the geometric area to get it...

28 January 2019 4,796 3 View

Should there be a correlation between band size on gel and SYBR green signal in the real time PCR run?

19 January 2016 4,703 3 View

Is there logic behind using Annexin V FACS with another antibody?

02 December 2015 605 3 View

High miniprep yield followed by extremely low Ct values?

I have been using Clone jet PCR cloning kit with a pJET1.2 blunt as a vector. The insert is from a clean PCR product that went through a blunting reaction (supplied with the cloning kit). The E....

19 October 2015 8,587 6 View

How do I use two way ANOVA on real time PCR results?

09 March 2015 8,424 6 View

How do I calculate standard errors for normalized log2 converted gene expression data?

I ran an experiment with a ref gene (3 biological repeats x 3 technical repeats) and normalized gene expression of my target genes to the ref gene. I decided to present the data as log2 fold...

05 January 2015 6,166 6 View

Has anyone had any experience with storing cell culture slides for a TUNEL assay?

22 December 2014 3,895 2 View

Can anyone recommend a process for preparing and storing slides for a TUNEL assay?

29 September 2014 8,964 2 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 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

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Does anyone have any ideas why drug-resistant malaria is increasing in Southeast Asia even when those drugs are no longer used?

When dealing with bacterial toxins, there are three varieties of microbes that develop in your petri dish: toxic, resistant, and susceptible. Initially, there are only toxic and vulnerable...

21 February 2021 6,084 5 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 3 View