Nested PCR || Principle and usage

Many many thanks, I appeals to keep it continue.

I start with thumbs up 👍 before I finish each of your videos !

Good work sir

please make a video on tailed PCR. thank u.

Some minutes and the topic is well understood... Thank you❤

Very well explained with diagrams and under 3 mins..

More than thank yoooooooooooooooooooooooooooooooooooooooooooou very much .. simple , to the point and GREAT!

Why don't we use secondary primer(specific primer) in the first place?

Your the best sir👌

Thank you

Wow

what if we design specific primer for specific region(that RED region in your video)??

boh ejas slide boh

Great

Perhaps a silly question, but do you combine both sets of primers in a singular master mix? Or do you add the result after a first PCR run with the second primerset seperately for a second run?

Thank you so much!

Thank sir ji

Amazing... explained within 3 min.. better than somu who would take 10min

Do you need to isolate your pcr priduct after first run before you run 2nd pcr? Or do it in same reaction?

Make a video on loop mediated pcr

Excellent video, but I don't really understand why having two sets of primers minimizes the probability that the second set of primers will bind non-specifically to other regions...

are you saying the first primer set, the green pair, acts independent of the 2nd pair? otherwise how are you eliminating nonspecific apmlicons?

am i to understand that the green primer set is depleted relative to red set of primers leading to more specificity of amplicons?

What ia the rationale of nested PCR?

هناك شرح بلعربي.

Excellent

What not we use 2nd set of primers first ?

I still don't understand because i thought a Primer amplifies everything from the binding site to the end of the molecule?

to clarify the doubts most people have in the comment section, the first set of primers are added along with the dntps ,buffer , taq polymerase first , then after the first round of amplification, the product from the first PCR reaction is used as the template for the second reaction, where the second set of primers are added.

thank you for this!

No this is wrong explanation

How do they filter out the non specific amplicons that are formed in the 1st round? They are not destroyed they stay there in the final whatever material is obtained.