How can I make a stable Mouse embryonic fibroblasts using GFP tagged for complex III of mitochondria please ?

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How do you assign allele numbers and sequence type after amplifying housekeeping genes by PCR for MLST?

How to assign allele numbers and sequence type after amplifying housekeeping genes by PCR for genotyping of S. pneumoniae strains by MLST using bioedit ?

19 July 2024 5,239 2 View

Sanger sequencing problems?

Hello, I have problems with PCR fragment sequencing (Sanger sequencing on SeqStudio). We perform sequencing reaction with BigDye™ Terminator v3.1 with subsequent purification with BigDye™...

29 April 2024 4,613 3 View

Need help getting rid of a persistent Peak on HPLC with PDA detector?

Hi, I am a graduate student using a Shimadzu HPLC with a PDA detector. This instrument was newly installed. Only the tech at installation and myself have run samples. I am using it for...

17 March 2024 5,352 2 View

What type of graph is this?

So, this may be a silly question, but what kind of graph is this? (see attached file). It seems to be a combo of a bar graph and a scatter plot? Can these be generated in Excel? If not, how are...

29 October 2023 468 0 View

Cytosolic / Nuclear Fractioning Inconsistencies ?

Hello all, I have recently started working with cytosolic/nuclear fractioning of HEK293T cells. My first few attempts were unsuccessful, in that there seemed to be pretty much equal...

25 June 2023 9,996 0 View

What is the prevalence of workplace violence?

the answer should contain globally, for Nigeria and for enugu state

22 June 2023 2,839 0 View

FLAG bead competitive elution?

Hello all, first post on ResearchGate! I have been performing immunoprecipitations with FLAG beads (abcam, ab270704), eluting using SDS loading buffer (with DTT) and boiling. The eluted products...

22 May 2023 9,054 5 View

Is the Fermi level of intrinsic semiconductor permanently changed to p or n type semiconductor by substrate influence? Why is that?

We know that intrinsic semiconductors will become p-type or n-type semiconductors by doping, will the work function of the substrate have the same effect on intrinsic semiconductors? This...

13 May 2023 149 1 View

10 point likert scale statistical analysis?

Hi, I’m struggling to decide which analysis I must do for my 10 point Likert scale questionnaire. Participants were asked to rate 12 classroom behaviours on their perceived levels of disruption...

29 April 2023 6,732 7 View

How can we increase motivation and decrease anxiety in foreign language courses speaking assignments?

How can we increase motivation and decrease anxiety in foreign language courses speaking assignments for high school students that are virtual remote learners?

09 April 2023 8,933 3 View

How can I prepare virus for a TEM or SEM imaging?

I have virus (viral hemorrhagic septicemia virus) in suspension and the experiment will not involve cells. What level of TCID50 is preferred?

11 August 2024 3,115 1 View

Which type of compound does lamda max of 218 indicate in a uv-vis spectrum of a partially purified compound through column and TLC?

A crude extract of fungal culture using EtOH was subjected to column and TLC and partially purified compound was obtained. UV vis spectrum of the compound/s has max absorbance at 218nm. The...

11 August 2024 9,801 2 View

1. If I can quantize the atom using this hyperbolic spiral and classical physics, could nature do the same?

If we map as a continuous motion an ionising electron (beginning its journey at n=1) in an H atom, a specific hyperbolic spiral appears (see animation). When we solve this spiral formula, we find...

07 August 2024 5,343 2 View

Is there anyone with experience in TEM analysis who can assist with a manuscript for an upcoming journal?

Hello dear colleagues, We have prepared a manuscript on NiTi-based alloys and are seeking a second opinion on our current TEM results. If you are a Ph.D. holder with experience in TEM and have...

07 August 2024 9,563 0 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

Could dyes amplify the spectrum of light to a specific wavelength?

I am interested to know the behavior of dyes toward light. Specifically, Blue dyes re-emit the spectrum, especially from the green zone (known as principal in LED lamps, and blue dyes are known...

05 August 2024 3,290 1 View

Which distribution type should I use when calculating the average particle size from TEM image? and how to calculate the error ?

average particle size calculation from TEM

04 August 2024 2,921 1 View

Why reactivity isn't increased with more empty spots in valence shell?

If from a geometric perspective the non-halogens, non-noble gases have more empty spots in their valence shell, and the filling/exiting of any of the empty spots in the shell constitutes a...

03 August 2024 4,787 2 View

Why is the molecule's orientation with an electric field affect polarizability?

Why is the molecule's orientation with an electric field affect polarizability? Electrons are diffuse enough to be independent with respect to orientation and effect of electric field on...

03 August 2024 7,843 1 View

Why doesn't chromium 2+ ion use all its d-orbitals to receive lone pairs from six waters in [Cr(H20)6)]3+?

I'm guessing it's because the ligand experiences too much electron repulsion or proton repulsion from the chromium to insert them close to the 3d-orbitals which are close to the metal nucleus. Is...

03 August 2024 1,370 1 View

Pavitra Ramdas

For transient transfection, I believe electroporation gives good efficiency in MEF. Reagents like Lipofectamine may give you around 10-20 percent transfection. However, lentiviral transduction is a good option for generating stable cell lines.

Lydia Amari

Thank you. Yes it will be stable transfection...

Praveen Joseph Susaimanickam

Hi Lydia. For making stable cell lines you can make use of the TALEN plasmids that introduce your gene of interest in the AAVS1 locus.

If you are new to that just have a look at the following link.

https://www.genecopoeia.com/wp-content/uploads/2014/09/Human+AAVS1+safe+harbor+User+Manual.pdf