Linearization of pPic9 plasmid using SacI?

More Thomson Patrick Joseph's questions See All

What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

23 February 2021 8,609 3 View

Describe a serious ethical dilemma you have experienced. Use the 12 questions developed by Laura Nash to offer a solution to the problem?

Describe a serious ethical dilemma you have experienced. Use the 12 questions developed by Laura Nash to offer a solution to the problem.

21 February 2021 9,133 2 View

How to perform Haldane correction on SPSS?

I was wondering if anyone knew how to perform the Haldane correction for SPSS (this is the definition of the correction -...

21 February 2021 2,882 1 View

Is it possible to quantify recombinant protein in the supernatant using SDS PAGE ?

I would like to quantify a recombinant protein (with native signal sequence) secreted by Pichia pastoris using the SDS PAGE. Since my standard protein (plant origin) is not pure, I would like to...

07 February 2021 3,880 9 View

How to analyze dendritic spines from tissue (Prelimbic cortex)?

Hi there, I am trying to measure dendritic spines from mouse fixed tissue, specifically, I am looking at the prefrontal cortex. I injected the PrL with a lentivirus to knock-down my protein of...

27 January 2021 9,251 2 View

High throughput screen to identify drug induced cell death mechanism?

So I am working with a drug that we know induces cell death in MDA-MB-231 cells. In other cell lines we have been able to identify that this drug induces ferroptosis. I have also done experiment's...

25 January 2021 3,587 3 View

How can I carry out analysis of pulp and paper properties according to Tappi standard method in the Lab?

Properties such as relative humidity (T 402sp-03), brightness and yellowness (T-452 om-02), whiteness (T-560 pm-96), opacity and absorption coefficient (T-425 om-01), tensile, breaking length and...

24 January 2021 308 3 View

PROCESS V3.5.2 macro for SPSS variable name length limitations?

I just installed PROCESS V3.5 in SPSS and am receiving the follow error when I try to run a model: ERROR: Variable names should be no more than eight characters in length. I don't recall...

11 January 2021 3,448 4 View

What is the kinetics that Cr6 on the surface of the soil will preferably exchange with the charge on the surface of the iron oxide nanoparticle?

I am researching the application of iron oxide nanoparticle in the removal of heavy metal (Cr6) from soil

10 January 2021 3,167 3 View

Can anyone direct me to where I can find the X-Ray elastic constants for WC-Co or just WC?

I have searched all over the internet and I seem not to be arriving at its value. I need this value to perform some XRD measurements. Any help would be greatly appreciated.

05 January 2021 916 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

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 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