How can I clone and express 3Kbp of DNA?

More Ganesh Shastry's questions See All

How is DNA labelled with radioactive sulphur? What are the advantages of this method over radiolabelling with phosphorous?

Labelling of DNA with radioactive sulphur

07 August 2014 2,504 3 View

Multiple ligands in autodock?

Can I dock multiple ligands in Autodock (either sequentially or simultaneously)?

07 August 2014 7,969 7 View

phage Infection

What are the Biochemical changes observed during Phage infecting bacteria ? One of them is membrane potential is anything else in the list ?? Thanks in Advance, Ganesh Shastry H M

06 July 2014 1,659 0 View

Can anyone tell me about the force involved in DNA ejection in bacteriophages?

I tried to understand both The Continuum Mechanics Model and Hydrodynamic model for DNA ejection but I cannot able to draw major differences other than the osmosis involved in hydrodynamic model....

06 July 2014 3,500 2 View

Why T4 bacteriophage tail protein in SDS resistant ?

T4 bacteriophage.

04 May 2014 9,854 0 View

Is there any assay to detect the T4 bacteriophage ghost in the given sample other than Electron microscopy?

Detection of T4 bacteriophage ghosts

02 March 2014 1,014 0 View

What is the best minimal media for E.coli strain B 11303(ATCC)?

I have to grow E.coli in a minimal media but I don't have M9 salt to prepare minimal media. Is there any other way to make minimal media for this particular strain? Thanks in advance.

02 March 2014 8,738 4 View

Is there any other way for the preparation of T4 bacteriophage ghost other than osmotic shock technique ?

Searching for any other technique for T4 bacteriophage isolation other than orthodox osmotic shock treatment.

01 February 2014 4,860 3 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Does crude extraction using NaOH and Tris work well with Fungi?

I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you

08 August 2024 4,733 2 View

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 View

Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...

06 August 2024 1,495 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Toshisangba Longkumer

What actually is your problem? Is it that you can't insert in a particular vector or you don't know which vector to use?

Ganesh Shastry

I have to express a viral gene which is of 3Kbp in E.coli I don't know which plasmid to use because the I have never worked with such a big insert