Why do I get my untreated pBR322 bands at between 2500-3000 bp but not 4000 bp?

I've conducted a quality check of my pBR322 using agarose gel electrophoresis. I got my supercoiled form plasmid band at around 2500-3000 bp and open circular form band at around 6000 bp. Why...

21 December 2016 9,261 3 View

Why do I get my untreated pBR322 bands at between 2500-3000 bp but not 4000 bp?

I've conducted a quality check of my pBR322 using agarose gel electrophoresis. I got my supercoiled form plasmid band at around 2500-3000 bp and open circular form band at around 6000 bp. Why...

11 December 2016 9,821 5 View

Oxidised phenol for phenol:chloroform:isoamyl alcohol method of DNA extraction?

I'm gonna carry out genomic DNA extraction from E. coli using PCI method. Is it still acceptable to use PCI with its bottom layer that has already turned slightly orange in colour? Will I still...

10 November 2016 5,980 3 View

Is there any other method that is able to provide yield as good as phenol:chloroform extraction method for genomic DNA isolation from E. coli?

I'm looking for an alternative method for the extraction of genomic DNA from E. coli. Is there a simpler way other than using phenol:chloroform and extraction kit? I've found a method that was...

10 November 2016 9,429 4 View

Will the concentration of SDS and Proteinase K affect the yield of DNA in PCI method of DNA extraction?

10 November 2016 636 6 View

Rationale of using pBR322 in gel retardation assay?

I'm currently studying the interaction of a synthetic antimicrobial peptide with DNA using gel retardation assay. I was given pBR322 as the study material for DNA. I'm currently looking for the...

09 October 2016 6,063 0 View

How to determine titre of increasing peptide concentration in gel retardation assay?

I'm studying the interaction between DNA (pBR322 plasmid) and a synthetic antibacterial peptide using gel retardation assay/electrophoretic mobility shift assay (EMSA). The plasmid is set fixed at...

09 October 2016 531 1 View

Hi all. Is there a way to conduct gel retardation assay without manually isolating plasmids from a bacteria?

The study is currently working on the binding properties of antibacterial peptide to plasmid DNA of Pseudomonas aeruginosa after the discovery that this particular ABP (known to have membrane...

07 August 2016 9,205 1 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 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

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

How to check the uv-vis absorption of nanoparticle by surface plasmon resonance?

I try to check the uv-vis absorption of Indium nanoparticle. Now I am measuring in solution, can I measure it in the form of a film?

01 March 2021 930 3 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