Is it harmful to breed for Cre Homozygosity initially?

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...

24 February 2021 5,759 3 View

Does anyone has a protocol for transfecting AML12 cells?

I am optimizing a CRISPR/CAS9 system in vitro before using it in vivo. I would like to transfect AML12 cells with my pAAV-sgRNA-vector to validate editing efficiency before generating virus.

26 January 2021 8,733 2 View

The impact of Methylisothiazolinone on haemoglobin quantification?

Hello everyone, I am a researcher working on a project aiming to develop an assay for Haemoglobin quantification. In our assay, prior to HGB quantification we dilute the sample in a dilution...

20 November 2020 7,316 2 View

Ansys Workbench Project view shows "Solved" but mechanical shows "Solve Required". How to Fix this without resolving ?

Hello everyone I am running a thermo-mechanical simulation on a Linux Cluster. In my Project view all the component systems seem to have solved. There is a file.rst file and and solver.out...

14 October 2020 2,631 3 View

Standard guitar tuning has two distinguished vectors (0 5 5 5 4 5) and (0 5 10 15 19 24). What is required to show they are tangent-cotangent bundles?

The pitch value set in music and guitar group in tablature are connected by adjunction of tangent-cotangent bundles. Tuning g is the tangent gradient to the flow of pitch on guitar. It determines...

04 April 2020 2,681 15 View

Can you ask for user input on pymol?

I want to write a script that asks the user to enter the PDB file so that the script can be entered (it prompts the user to enter it), the user enters the 4 character code, then the rest of the...

23 February 2020 1,184 1 View

Why cani I not find Constantino Manuel Torresd on Research Gate Search Enginge?

I just read a new paper posted here by Constantino Manuel Torres. However when I put his name in the search engine here, it says there is now one here with that name? I wanted to send him a...

13 July 2019 2,496 3 View

How do I design primers specific to transfected plasmid and not genomic DNA?

I recently introduced a Lenti-Puro Guide vector containing a specific sgRNA sequence into a cell line. I would like to sequence the ddCas9-sgRNA cell lines to confirm that each sgRNA is accurate.

04 February 2019 6,681 3 View

How to make culture media more basic (not too basic) without a pH meter or strips?

Hi, A friend of mine suggested that the reaction that makes Salmonella colonies appear black on XLD or TSI agar might show an increased rate at a slightly lower pH. I was thinking of adding a bit...

24 July 2018 2,916 7 View

Does anyone has protocol for p53 IHC in frozen sections?

I need to detect p53 in murine E15.5 embryos that overexpress this protein. The embryos were collected and fixed overnight in 10% Neutral Buffered Formalin. Then, I cryopreserved these embryos in...

23 May 2018 2,592 2 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

Are there lab markers/pens resistant to Roti-Histol?

Hi, I'm looking for a pen/marker to write on the smooth, transparent (non-frosted) part of microscopy slides which does not get washed away by alcohols and most importantly Roti-Histol. I want to...

24 February 2021 9,902 5 View