Has anyone tried to scan fluorescence IHC slides with microarray scanner?

I have an old slide microarray scanner (VIDAR Revolution 4200) and would like to give it a "second life", as IHC scanner array (fluorescence dyes), FISH over tissue or similar. Has anyone tried...

24 February 2021 9,210 1 View

Different antibodies show signal at same MW in SDS-PAGE, what is wrong?

16 February 2021 2,857 9 View

The modern system of international relations: unipolar or multipolar world?

Do you think that there is one center of power in the modern world, or are all countries equal in the international arena?

01 January 2021 7,026 2 View

Mie scattering of a Gaussian with curved wavefront by a sphere?

Can anyone please suggest some references? I would like to simulate scattering of a fundamental mode of a fiber by a microsphere. I am only familiar with a plane wave case, but in case of a curved...

01 January 2021 1,440 3 View

Blood and tissue DNeasy mini kit 260/230 contamination?

I've been trying to extract DNA from rat liver tissue using Blood and tissue DNeasy mini kit. I followed handbook procedure and all of my samples had low 260/230 (

21 December 2020 7,100 3 View

Student's t-test or DeSeq2 for RNA-Seq data analysis?

Hi everybody, I am analysing RNA-Seq data of patient samples. One group of samples has a certain monoallelic deletion, the other group 2 copies of the wildtype gene. Now, I want to compare the...

28 November 2020 5,977 3 View

Cre-loxp system does not work (lines #017455 and #017320, JAX). Why?

We bought this year two lines (#017455 and #017320) to get specific expression of channelrhodopsin2 (ChETA, in this case) in parvalbumin-positive (PV) interneurons. We hoped that crossing a mouse...

16 November 2020 8,701 1 View

Why after cloning recombinant pET in E. coli DH5a I get a very low plasmid yield after extraction?

I chemically transformed a ligation product (pET28a and my gene of 3051 bp) into E coli DH5alpha competent cells. I plated 200 and 800 ml of my transformation on plates of LB agar Kan (25 mg/ml)....

13 September 2020 4,421 3 View

Good markers to distinguish D1R/D2R striatal medium spiny neurons?

Hi! I want to distinguish D1R and D2R subpopulations of medium spiny neurons in striatum in immunofluorescence. Most of the papers I found use transgenic EGFP mice targeting this receptors, but I...

31 August 2020 3,863 2 View

does anyone have the user manual of the geostrain program?

Hello, I am looking for the user manual of the geostrain software, I have downloaded the software but the user manual is not found. I will be very grateful for your help Cheers

14 June 2020 4,169 1 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 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

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

Hi i want to ask that in order to find out LC 50 (96 hours) of particular toxin for fish. Should we keep the animals with food or without food?

if we expose the fish after acclimatization to particular toxin in order to find out LC 50 (96 hours).Should we keep the fish off feed or with feed for that duration. Some literature show that...

24 February 2021 357 2 View