How temperature affect plasmid DNA conformation?

I conduct a thermal stability test for plasmid DNA. In which I put plasmid DNA under 65-degree celsius for about one week. After that, I run an agarose gel check. Here below is what I got: 3 left...

11 December 2016 8,159 4 View

What is the reason for the complete loss of transformation efficiency?

I have conducted a test on transformation efficiency of circular plasmid DNA after some harsh storage period (in which I put the plasmid DNA in 60 Celcius degree for 2 weeks). And results showed...

09 October 2016 3,275 4 View

Linearized and Circular Plasmid DNA, Which is more stable under same storage condition?

I would like to ask if someone has any experience or Ref on the stability comparison of the above said plasmid DNA (linearized and circular plasmid DNA, stored at same storage conditions). What...

09 October 2016 9,270 6 View

What are the methods for nucleotide ratio quantitation?

Providing that I have a single stranded DNA (short fragment ~30 nt) with unknown sequence. What are the experimental methods which could help to determine nucleotide ratio (percentage of A,T,G,C)...

03 April 2016 314 2 View

How to measure adsorption of biomolecules in liquid on solid surface?

03 April 2016 2,400 3 View

What is the problem with my bacterial cell growth in LB media?

02 March 2016 3,904 7 View

What universal primer should I choose for plasmid sequencing?

11 December 2015 2,223 6 View

What are the differences between Nanoparticles and Nanopowder?

I am in need of using Manganese nanoparticles but not quite familiar with the synthesis. There is nanopowder of manganese oxide commercially available. In this case, could I disperse nanopowder...

10 November 2015 3,996 5 View

Could an incorrect dilution affect the reaction outcome?

I think there is a difference between these 2 dillution methods and therefore it might affects the reaction (A+B=AB). Could you please give me some clear explainations on this: I have: Stock A: 10...

31 December 2014 2,909 3 View

EcoR I does not fully digest my DNA. Why?

I used a 80 bp ds DNA which contains EcoR I recognition site in the middle of the strain, my objective is to obtain 40 bp ds DNA fragment. I used the protocol supplied with my enzyme (from...

07 August 2014 7,337 8 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

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 provide the Support vector regression code structure of LIBSVM MATLAB or python ?

Please, if Gaussian Parameter optimization is possible in this code structure

28 February 2021 278 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

Does anyone have a sequence for the human CD34 promoter?

I'm looking for the human CD34 promoter sequence to engineer into a vector construct. Can anyone recommend a good source for this sequence? Thanks very much, Laura

25 February 2021 8,205 1 View