Hello.. what is the best plasmid that can be used recombinant gene extraction for pseudomonas aeruginosa?

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High Power control for 48V battery pack - High current control?

Hi, I am working on 48V Lithium-Ion battery pack. I am wondering if there is fast MOSFET (or any other technology), to control the current of charging and discharging the battery with high...

20 January 2021 1,717 4 View

GROMACS: Cannot rename checkpoint file; maybe you are out of disk space?

Hi! i am running a simulation using a cluster and I have encounter an error during my nvt equilibration like this: Cannot rename checkpoint file; maybe you are out of disk space? I deleted some...

03 January 2021 3,544 2 View

How can the resistance of paving asphalt be increased to acid rain?

Modification of the rheological properties of asphalt

25 December 2020 7,390 2 View

Can someone please share the matlab code of FBMC/OQAM PAPR reduction with TR and PTS methods?

I am trying to work on reducing the PAPR of the FBMC/OQAM with the TR and PTS methods. for that if someone can share with me one of these matlab codes. my e-mail: [email protected]

19 December 2020 7,764 1 View

Appreciate the quality qualities of cheese ?

How to estimate the acidity of a cheese?

13 December 2020 4,890 4 View

What is the appropriate polymerase to amplify gDNA greater than 10kb?

I am going to clone gDNA fragment > 10 kb into a pCR TOPO vector. I performed the PCR using KOD PLUS Neo polymerase for 3-steps PCR reaction, I am not able to find the specific size. Several an...

03 December 2020 1,939 5 View

How to get the data of the changing the distance between two Pionts with the applied force?

does anybody knows how can i get the change in the distance between the notch nodes (two points showing in the figure ) vs the applied force in every step as a table out put or curve in...

23 November 2020 352 3 View

Why I got low CI in the EBSD?

Hi I got a low confidence index in the EBSED analysis even after days of polishing using 1 to 0.02 um suspension using 1 lb force, the sample was Al/W composite (has intermetsllic interface)...

26 August 2020 3,547 4 View

Hi Is it needed to etch a sample in order to see dislocations? SEM or TEM microscope should be used for that?

hi please I have Al/W sample and I am looking for screw dislocations on the intermetallic surface, my question is: Should the sample be etched? and should I use the SEM or TEM microscopy? what...

06 August 2020 9,703 2 View

How can i increase my validation and test accuracy?

I am using google colab to train a model for sign language recognition using this dataset: https://www.kaggle.com/feronial/turkish-sign-languagefinger-spelling I tried different architecture...

08 May 2020 4,098 5 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

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