Can be episomal plasmid vector amplified in mitotic cells?

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Which AAV serotype is the best for SY5Y and HT22 cells?

I am not sure; is AAV2 best for SY5Y and HT22? Is there any comparative study for these lines? Also, Can i use same serotype vector for the two cell lines?

02 October 2020 3,328 2 View

Can i use ORI in my plasmid vector for replicating it in SH-SY5Y cells?

I want to use ORI in my plasmid vector for replicating it in SH-SY5Y cells like in 293E or 293T cells with SV40? Is there any ORI that can be replicated in SY5Y cells?

24 September 2020 2,541 3 View

How do you dissolve and inject indomethacin for rat studies?

I dissolved indomethacin (Santa Cruz Biotech - SC-200503) in DMSO and prepared stock of 25 mg/mL In my study, I inject 10 mg/kg i.p., injection volume 0,5 mL and I work with 250-300 g rats. When...

13 September 2020 5,659 3 View

Why do i get wrong total pressure data from Ansys Fluent?

Hi everyone, I have been working on a laminar compressible microtube flow analysis (supercritical fluid). Mass flow inlet, pressure outlet and constant wall heat flux boundary conditions are...

17 December 2019 1,403 4 View

How can cyclic symmetry be defined in dynamic, implicit analyse in Abaqus?

Hi, I need some support to define cyclic symmetry in dynamic, implicit analyse. As I have found out that cyclic symmetry under interactions is only usable in static, visco, frequency, steady...

26 November 2018 3,020 5 View

How do I find references to the size of the cDNA sequence I isolated?

12 March 2018 2,101 3 View

Can be possible the dna binding domain's binding to two or more different DNA sequence in different enhancers?

Can be possible the transcription factor's dna binding domain's binding to two or more different DNA sequence in different(or also same) enhancers? Or there is no exception, it must be specific...

23 February 2018 8,240 3 View

Why is ETOTAL negative in ABAQUS Quasi Static Analyse?

Hi, I did an quasi static analyse. As a result of my analysis, total energy of the output set (etotal) is everytime negative and after some steps etotal increases exponentially in negative area....

23 January 2018 7,094 6 View

How can I find out the rotational stiffness of the bearing with static structural analysis?

Hi, I have some problem about the rotational stiffness of the bushing. Actually, I would like to change the bearing balls with bushing joints in ANSYS Workbench. I have a normal bearing with the...

01 November 2016 7,088 3 View

How can I remove orientation node from beam element in ANSYS WB?

Hi, I have a problem with orientation nodes of beam element in ANSYS. My Structure: - 3D Structure (Every node have 6 DOF) - 12 Beam Elements - 13 Nodes - 2 Fixed supports at the edges - After...

24 February 2016 541 3 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

What solvent can I used for Paal Knorr reaction between 6-amino hexanoic acid and 2.5 hexanedione?

I am try to make the Paal Knorr reaction between 2.5 hexnedione and 6-amino hexanoic acid, my problem is type of solvent to use, because 6-hexaminohexanoic acid is soluble in water but I am not...

02 March 2021 4,443 2 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

Can anyone share his experience regards Smoothing in origin?

I am working on TiO2 composite material and the results of XRD have a lot of small/junk peaks and while I apply a smooth filter in Origin then the other characteristics peaks disappear so anyone...

26 February 2021 8,183 3 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

Michael Halim Popular answer

Interesting question. Following the discussion.

Walid Hassene Hamri

Hi,

You may find what you looking for in the link below:

Article Episomal vectors for gene expression in mammalian cells

John Schloendorn

This can often work. For example, CHO cell lines for transient expression are made in this way.

Michael Halim