Does anyone know a protocol to remove a derivative of a mini-F plasmid from an E. coli strain in order to use the resulting strain as a host?

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Is there any tool to measure impact of climate change on wellbeing of people of a particular community? Please suggest?

Climate change is a topic that should be given so much importance. How can I go forward with this topic for studying the impact of climate change on people's well being?

14 September 2020 3,141 4 View

How can I measure the impact of climate change on the wellbeing in India?

I am very much curious to learn the impact of climate change on the psychological wellbeing or wellbeing of the person in general. Is there any standardized tools to find this out. How can I go...

29 July 2020 1,141 4 View

Normality Test problematic: too low samples in ANOVA. Any solution?

Dear All, An experiment was carried out, using as factors the temperature, the humidity and in addition, it was carried out for 6 different species. The total sample number of species analyzed is...

13 December 2017 577 3 View

Should we use different frailty measurement tools in different settings? Or just one measurement tool for all settings?

There are many frailty measurement tools used in hospitals and in primary care services. Should we use different frailty measurement tools across the different settings, or should we just use one...

29 June 2017 2,688 2 View

Are you interested to participate in this project as a postdoctoral researcher?

We opened a position at the Department of Civil Engineering of the University of Porto for a postdoctoral researcher with expertise in Bridge Monitoring/ Damage detection/ Structural Assessment....

01 January 1970 7,395 32 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...

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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...

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How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

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Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

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24 February 2021 6,310 3 View

Best plasmid could be used as a standard curve for AAV titration?

I have been trying to have a good AAV titration for my production and I have tried different plasmids to set up as a standard curve but each one behaves differently. i am trying to use ITR for...

23 February 2021 5,167 3 View

Alejandro Martin

You can do plasmid curing with acridine orange or ethidium bromide. Essentially, you grow the plasmid-bearing cells on the presence of sub-inhibitory concentrations of either compound, plate and then screen the resulting colonies for plasmid loss. The process is not exactly "efficient" though; in a best-case scenario at most 5% of the screened colonies will have lost the plasmid.

Hope it helps!