Bacterial DNA Extraction

#DRMAKKY #microbiology #labtechniques #lifescienceskills
In this video, we need to explain How to extract the bacterial DNA using TE buffer pH 8.0 including DNA precipitation and purity.
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[@wanlola1691]

Very good lab

[@DRMAKKY]

thanks

[@alinoh3564]

Nice

[@DRMAKKY]

Thanks

[@kinzlala8259]

Good job

[@DRMAKKY]

Thanks

[@nadamohd4645]

Thanks

[@DRMAKKY]

Welcome

[@FHNSTU]

Great job, Professor. Thank you for your contribution to the world.

[@razalilee2688]

Thanks

[@DRMAKKY]

Many thanks for your comment

[@nathanomambia6085]

good job doctor ,happy to hear you from kenya,,am a student of biomedical science and am really helped

[@DRMAKKY]

Many thanks Nathan Omambia .. I hope to follow and subscribe to get my updated videos

[@A4arooj]

This was so far an amazing video .

[@drvikasgautam6859]

seems innovative. The purity of DNA is good, but the concentration is less. I will also try this method guys.

[@DRMAKKY]

Thanks for the comment but in our video the concentration is good. If you got low concentration please check the extraction handling method and the purity of your organism.

[@larawan4598]

Good job dr

[@DRMAKKY]

Thank you 🙂

[@dashingbadar5004]

Great effort👍

[@DRMAKKY]

Thank you! Cheers!

[@SaumyaSharma123]

Hello sir
I have following questions-
1. What we call this technique that you have used ??
2. What if I have -20 degree maintenance in a lab deep freezer ?? Can I put eppendorf in it?? Any alternative?
3- Can I use this technique to extract DNA from microspodian spores ?? It is a sister fungus group that resides in the abdomen of bees..
I really need assistance. Plz help me out

[@DRMAKKY]

Hi Saumya and thanks for ur questions .. For Q1: this is called a simple technique to extract bacterial DNA using TE buffer. For Q2: actually -20 u can use but u need to increase the time, on the other hand, u may facing NOT accurate extraction. So, u can try -20 otherwise u have to use -80. For Q3: NO this technique is for bacteria. If u need to extract the DNA for fungus u need to use an extraction kit. I hope it was beneficial .. Good luck & Thanks again

[@SaumyaSharma123]

@@DRMAKKY thanks sir. Means a lot. In support of your third answer, I want to tell u that I have used DNeasy plant mini kit six times but I didn’t get results yet😞 on electrophoresis

[@DRMAKKY]

@@SaumyaSharma123 I think this is due to u r not using DNA fungal extraction kit coz if I'm not mistaken the DNeasy plant mini kit for plant DNA extraction. Try to change the method (kit).

[@jedeiahserondo2860]

hello sir! i have a question,
1. Explain the purpose of each step in the protocol. How will each reagent interact
with the biomolecules in the cell ensuring the isolation of genomic DNA?
2. Aside from organic extraction, what other methods can be used to extract DNA
from cells?

[@DRMAKKY]

Hi .. thanks for ur questions. For Q1, u need to check the DNA concentration & purity using microvolume spectrophotometers. For Q2, ther're many other methods using different Extraction Kits.

[@mohamedaliabdelbaseer1311]

تحياتى ليك د عصام. محمد عبد البصير جامعة الازهر

[@samridhipushkarna4162]

Hello sir
Can we use the plates (inoculated and already kept in refrigerator for past 2-4 days) for isolation by this method? Or should the plates also be inoculated again for fresh batch?
Second, if we have -80, for how long should we do that freezing step?

[@DRMAKKY]

Thanks for ur good questions: #1 this depends on the growth rate of ur bacteria (slow growth rate u can use it for the past 2-4 days), if fast growth Ex. within 12-18hra better to get fresh culture); #2 in such case u can try for only 30 min OR 1hr for best results.

[@samridhipushkarna4162]

Also sir, can you please post another picture of the DNA ring that you mentioned.

[@DRMAKKY]

Sure I'll try to find a clear one .. Thanks

[@samridhipushkarna4162]

@@DRMAKKY sir, I tried this method but could not see any ring. Also my culture is from NB agar plate so is that creating it's difficult to see the ring? Since you are using McFarland.

[@samridhipushkarna4162]

@@DRMAKKY I am trying to isolate dna by other methods also but evey time I rna contamination. Is there any other way to remove rna contamination if we do not have rnase A

[@DRMAKKY]

​@@samridhipushkarna4162 I appreciate you asking. I believe you should try to increase the concentration of your bacterial suspension and/or double-check all of the reagents and extraction processes that were employed.

[@olachiosuagwu3882]

please can you do a video on PCR amplification method and 16s rRNA sequencing

[@DRMAKKY]

Many thanks for your comment and I'll try my best to do so soon 👌👌

[@achyuth7586]

1.my question sir why freezing is done
2. and why freezing and heating is repeated twices

[@nawandnamiq4072]

This principle same for all bacteria?

[@AbhishekKumar-yp7si]

13400'g' sir centrifuge is denoted by rpm , i little bit confusd

[@DRMAKKY]

@Abhishek .. good question, please check this link for details tools.thermofisher.com/content/sfs/brochures/TR0040-Centrifuge-speed.pdf

[@betatheta6611]

Sir, i have a question!
Can i use this method for rhizobial dna extraction?

[@DRMAKKY]

Yes, you can

[@easybiology9851]

If lysozyme is not being added in case of gram positive bacteria

[@BAjey-nb1ic]

Spirulina DNA isolation you putVoides sir

[@DRMAKKY]

No, I don't have videos for Spirulina in my Channel

[@junheng1258]

Can i use broth culture in this method？

[@DRMAKKY]

Thanks for your question .. but let me know which both cultures you are talking about?

[@junheng1258]

@@DRMAKKY nutrient broth culture, sir

[@junheng1258]

@@DRMAKKY I'm trying to identify unknown bacteria culture, i've done isolation and culture them in nutrient broth, if i want to proceed, do i need to subculture again from broth to plates, or i can just take the sample from broth

[@DRMAKKY]

@@junheng1258 Hi .. You can use any, but the most important is the culture should be freshly prepared (24h-48h aged)

[@junheng1258]

@@DRMAKKY Oh, okay, thank you for your reply sir!😃

[@PersefonaeParacelsus]

please clean up that microcentrifuge ToT

[@johnvet8522]

The 260/230 value is very low.

[@barbiedoll8322]

can i get your mail id , for internship opportunity in your lab , i will be highly grateful. CURRENTLY I AM A STUDENT OF INDIAN INSTITUTE OF SCIENCE EDUCATION AND RESEARCH .

[@fazlirasa2334]

thanks

[@DRMAKKY]

Welcome

[@NaturalGrower-dk8jg]

Good job