PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

Quantifying leaking DNA from bacteria - what's the secret?

Hi, all! I need to determine if my treatment causes membrane damage in bacteria (S. aureus and E. cloacae), but I cannot use a fluorescent probe such as PI and similar ones because my compound is...

24 February 2021 2,006 2 View

Please let me know how to remove mycoplasma contamination from cell line?

I am facing a problem in maintaining the cell line of HepG2 and HepG2.2.15. I have used plasmocin Tm treatment (Invivo gen) Cat.#ant-mpt Lot #MPT-41-01 for more than a month. I have also used...

21 February 2021 6,046 4 View

Western blot protein of interest falling at a lower weight than expected?

My protein of interest is DAGLa (115 kDa). We received the antibody for this from another researcher who reliably shows bands at this weight. We do see a light banding at this weight, but the...

11 February 2021 6,009 3 View

Which Journals would you advise to submit a Letter which briefly talks about neurosurgical treatment of neuro-oncological patient during Sars-COV-2?

Possibily not an open access journal which does not add any publishing costs once being accepted. Thank you in advance

09 February 2021 717 6 View

Sample Size for Quasi-Experimental Quants using one-tailed t-test?

Hi, sorry I'm stumped. I'm going to do a study on the leadership styles (using the MLQ Form 5X) of minority women administrators in medical schools. How will I calculate the sample size? Thank...

30 January 2021 2,865 4 View

Can you do Ponceau staining after loading control for Western Blot?

Hello everyone! I forgot to take a picture of my Ponceau stain prior to starting my first round of primary and secondary antibodies. I'm currently doing my loading control now and was wondering...

25 January 2021 757 3 View

What is the correct way to apply a Design effect for sample size calculation when the sampling method is linear systematic random sampling?

When we are doing sampling by means of simple systematic random sampling,how do we calculate the design effect for use during calculation of sample size for our work?

20 January 2021 5,328 3 View

Anybody to help me with Code source for DOUBLE CONCAVE FRICTION PENDULUM BEARINGS Please?

Hello, would you please share the codes for Base Isolation of structures using Double Concave Friction Pendulum Bearings? I am doing a related research but as I am just a beginner in MATLAB, I...

18 January 2021 6,683 1 View

How to generate .dlg files on PyRx?

Hi. I'm using the PyRx version 0.8 and getting the error message: WARNING: Filename mismatch: "2DFN_ADP_CL.maps.fld" :: "..\2DFN_ADP_CL.maps.fld" C:\Program Files (x86)\The Scripps Research...

17 January 2021 843 3 View

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 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

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Markers of mitochondrial activity in FFPE tissues?

Hello everybody, Does someone have any experience with assessing/extimating mitochondrial activity on FFPE tissues? Precisely, no TOM20, ATP5B, etc: we are trying to get an approximative idea of...

24 February 2021 4,618 4 View

Can a Golgi-stained dendritic spines picture processed by Image J be directly used for publication?

24 February 2021 3,348 2 View

Is there an appropriate CD8 antibody for IHC-P for staining human breast cancer tissues?

Hello, We are planning on staining Formalin-fixed paraffin breast cancer tissue sections (FFPE) with CD8 Ab via immunofluorescence (Tissue IF). Based on our existing protocol and imaging...

24 February 2021 9,578 4 View

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