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

Concentration of plasmid pBR322 to be used for DNA-antibacterial peptide interaction study?

09 October 2016 9,883 2 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

Looking for Materials with high thermal expansion coefficient and high temperature resistance?

03 March 2021 8,272 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

I am interested in medicinal chemistry field, which books are highly recommended to learn from?

I am at the beginner level in drug design and drug discovery, and I am interested to learn more about this field. Which books are highly recommended to start from and gives me all essential...

02 March 2021 8,437 2 View

How to grow E.coli on the low salt LB agar plate with 50 or 100ug/ml of Blasticidin?

Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...

01 March 2021 2,027 2 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 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

Are there lab markers/pens resistant to Roti-Histol?

Hi, I'm looking for a pen/marker to write on the smooth, transparent (non-frosted) part of microscopy slides which does not get washed away by alcohols and most importantly Roti-Histol. I want to...

24 February 2021 9,902 5 View

How to empty large intestine in mice ?

Hi all I am working on colon-targeted drug delivery for which I kept mice fasting for 6h using fasting grills during scotophase (dark period). I observed liquid fecal content in the cecum and...

23 February 2021 7,701 2 View