Reagents for histology slides?

Dear all, I am planning to study histology and want to do that at home. I have only stained a blood smear with Giemsa but now want to try a wider range of animal tissues. What reagents will I...

24 February 2021 5,430 3 View

What is the optimal propagation regime for pET-1M(KanR) and pMCSG7(AmpR) plasmids in DH5alpha E.coli?

Dear all, I am trying to harvest the low-expression plasmids in the mentioned E.coli strain. I constantly gel very low concentrations (30-50 ng/uL while the concentration is usually around 100...

08 February 2021 4,255 4 View

Does increasing antibiotic concentration also increase plasmid production in DH5a E.coli?

Dear all, I am propagating some plasmids with a human gene and an ampicillin resistance gene. My logic is that if ampicillin concentration will be increased, the concentration of plasmid (pSF)...

14 January 2021 779 3 View

Why didn't site-directed mutagenesis work?

Dear all, I was doing site-directed mutagenesis with one mutagenesis primer (fwd) and one normal primer (rev) to amplify a 6.5-kb plasmid. Then KLD was performed with dnp1 destroying the original...

10 January 2021 992 6 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

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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

LEDs on Vis Light photo degradation of MB water sol via N doped TiO2 in a mini flume?

Good day to all, I have no experience with light emitting diodes. I am building a glass mini flume to demonstrate the photo degradation of Methylene Blue in water running over self cleaning...

26 February 2021 4,026 5 View

Why do a weight decrease for thermal degradation in nitrogen gas condition?

Hi all The question is related to TGA(Thermogravimetric analysis). Thermal degradation easily understood but why do samples weight decrease? altough just binding is broked, the weight of...

26 February 2021 1,100 5 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 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

Steps to calculate activation energy by coats & redfern method?

I'm a chemistry student and I'm researching the kinetic degradation of mixed plastic waste. I'm having trouble calculating the activation energy by the coats & Redfern method. The data that...

24 February 2021 974 1 View