What is the best method for extraction of large plasmids?

More Mohamed Sebak's questions See All

Anyone can help please about the oxygen level of fermentation of actinobacteria in the bioreactor?

I'm asking about the optimum oxygenation level or range for bioreactor fermentation of some soil actinobacteria for production of natural products. Thanks in advance

02 March 2018 5,775 8 View

What is the best solvent used for antimicrobial activity screening other than acetone and ethyl acetate?

I have problem that ethyl acetate and acetone have some antimicrobial activity against some of my standards strains, so I shouldn’t use them for antimicrobial activity not to affect my results...

08 September 2017 7,194 4 View

What’s the best method for determining the MIC and MIC 50 of bacterial natural products against some bacterial standard strains?

I’d like to know how to prepare the standards strains broth in the best way and how to do the test and interpreter the results I know that it should be done in triplicates but I’m not sure about...

08 September 2017 5,479 12 View

Can soil actinomycetes isolates tolerate high temperatures like 42?

What's the highest temperature could actinomycetes isolates tolerate and what's maximum time for these high temperatures Please anyone help

06 July 2016 9,203 4 View

What is the optimum concentration of Genomic DNA and primers to get perfect PCR products?

I want to get sharp bands of PCR products on the gel electrophoresis without any tailings, primer dimers or non-specific products. What is the best concentration of both primers and DNA to get...

11 December 2015 9,748 7 View

What is the best antifungal used in isolation of a mix of bacteria and fungi?

I have some environmental samples that contain both fungi and bacteria and I want to isolate only bacteria What is the best way to do that and if I want to add antifungal in the media, what is the...

11 December 2014 1,584 10 View

What is the best method for identification of actinomycetes biochemically?

I had isolated some isolates suspected to be actinomycetes. What is the best method for identification of them biochemically? Please help if you can Best regards

09 October 2014 9,601 3 View

Can anyone help me to collect non-clinical microbial strains producing biologically active secondary metabolites?

If I want to have bacterial samples which can produce biologically active secondary metabolites, what is the best way to do that and what are the sources from which I can collect them? Please help...

09 October 2014 9,052 16 View

Does media selection affects the production of novel biologically active compounds from the microbes?

If I want to have bacterial samples producing novel biologically active secondary metabolites, what is the best way for selection of the media for cultivation of these bacteria to produce new...

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What is the best media for cultivation of actinomycetes to assist production of biologically active secondary metabolites?

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09 October 2014 6,463 9 View

Is there a problem with my RNA pellet?

Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...

11 August 2024 7,082 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...

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RNA Extraction Using Hot Borate Method No Longer Working?

I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...

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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

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

What precautions should be taken while handling S. aureus enterotoxin Type B in the lab?

I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...

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How do soil microflora interact with plant roots and influence plant nutrition, health, and productivity?

06 August 2024 9,618 3 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

Robert J Mitchell

Not certain if it is the best but I use the Qiagen kit (or similar kit) but when I elute, I heat the elution buffer up to about 45~60ºC. This improves the elution yield and the overall purification.

Ying-Tsong Chen

Qiagen kit will do. It works on plasmids sized from 100~200-kb in our lab (E. coli).

Claudine Etienne

What is the purpose for isolating the plasmids? Will the bacteria be studied, The plasmid, or both? What is the purpose of the study?

Mohamed Sebak

Purpose is studying of the plasmids

Shahida Hasnain

Gel electrophoresis of total cell lysate (Eckard method) then purify from gel using kit

Tanedjeu Kemgang Sonfack

Dear Mohamed, this paper attached will probably help you.

good luck.