RAPD doesn't need forward or reverse Primer. Only one primer will bind to the sequences complementary and fragments of different sizse will form. By running this PCR product one will see the variations among the sample in study. Rapd doesn't need a specific sequence.
a few queries that I have are as follows- 1) When we design primer in accordance with the sequence, how come its random? 2) During the initial part of the video, you said that the 'mutated' sequence will not allow the primer to bind properly, hence we get lighter bands of less concentration. But what if the mutation occurs upstream of the primer binding site? this would mean that the primer will bind properly and also we will also get a complete band because we will have reserves of all four dNTPs anyway. What will be the result then? 3) Wouldn't the SNP s during CAP encounter the same problem of primer binding while doing PCR ?
hey somu i am confused about this is whether we use( randomly designed primers) will be differ for one individual to another comparing individual or we use same ( randomly designed primers)for other comparing individual also ?
one basis problem that i found regarding the word Random....... here u told amplification should be random...... but in whole video u were talking about the designing of the primers according to the sequence information........ so my question is that... if process is random amplification then what is the need of designing of specific primers according to sequence information??? As per my knowledge in the method of RAPD we use random primers without the prior knowledge of sequence information....... please clarify this....... thank u...
This is a confusing process. Why did you sequence the target DNA you extracted from the gel and then still need to do a restriction digest? Surely the sequencing would give you the final answer as to whether or not there is a change, you could just align it?
During the initial part of the video, you said that the 'mutated' sequence will not allow the primer to bind properly, hence we get lighter bands of less concentration. But what if the mutation occurs upstream of the primer binding site? this would mean that the primer will bind properly and also we will also get a complete band because we will have reserves of all four dNTPs anyway. What will be the result then?
my question is that... if process is random amplification then what is the need of designing of specific primers according to sequence information??? As per my knowledge in the method of RAPD we use random primers without the prior knowledge of sequence information....... please clarify this....... thank u...
please tell if template DNA is mistakenly mixture of 2 genotypes but we only want to amplify one genotype... then it'll show mutation ???? or pcr product will not form ????
ek request hai, Sir aap apne poorane videos ko phir se nai videos daaliye sir in sab ya volume kam hoti hai aur user interface thoda dull hai. (Thank u for all your videos)
An interesting clip but I will argue strongly with your differentiation between polymorphism and mutation (from 6:50 to 7:50 min). From a general consensus polymorphism is a result of mutation!
Although I really like his videos and his way of explaining the things but in this case the information regarding primer is wrong. Kindly note that RAPD markers are DNA fragments from PCR amplification of random segments of genomic DNA with single primer of arbitrary nucleotide sequence.
Nooooo, RAPD doesn't use two primers, it only uses one and it is not a specific one, it's and arbitrary one that binds randomly, that's why it's called RANDOM. The amplification of DNA sequences coming from multiple individuals using the same primer can emphasize the differences after running the gel, the absence of a band in the gel might indicate a SNP in the region that the primer binds, because the primer was unable to bind to that region and thus it couldn't be amplified like in the case of the individual that lacks the SNP in the same region.
thank u for nice presentation. i have one some queries 1. why should the length of RAPD markers is 10 base pair and why the length of SSR markers is more than 20 base pair
hello Suman. it is really very informative.bt my question is here you took the example of same gene sequence of two individuals? i.e. one is normal and another is mutated ?
I hope you can help me more I have this project about SNP and microsatellite mainly the detection methods can we say this method in the video is a detection technique as TaqMan Snp and others. I hope you can help me all the best (3 days to my presentation ) 😢😦
Why did you design the primer? Aren't the primer supposed to be of random sequence? For random amplification?
Very good clarity in explanation. .
Thank you
RAPD doesn't need forward or reverse Primer. Only one primer will bind to the sequences complementary and fragments of different sizse will form. By running this PCR product one will see the variations among the sample in study. Rapd doesn't need a specific sequence.
a few queries that I have are as follows-
1) When we design primer in accordance with the sequence, how come its random?
2) During the initial part of the video, you said that the 'mutated' sequence will not allow the primer to bind properly, hence we get lighter bands of less concentration. But what if the mutation occurs upstream of the primer binding site? this would mean that the primer will bind properly and also we will also get a complete band because we will have reserves of all four dNTPs anyway. What will be the result then?
3) Wouldn't the SNP s during CAP encounter the same problem of primer binding while doing PCR ?
hey somu i am confused about this is whether we use( randomly designed primers) will be differ for one individual to another comparing individual or we use same ( randomly designed primers)for other comparing individual also ?
if it is random why do we need specifically designed primer?
RAPD do not involve restriction digestion
Prier sequence are designed randomly
Ur far more better than our faculties
Thank you so much for appreciating my efforts
Suman you are really genius ... Outstanding performance👌👌👌
one basis problem that i found regarding the word Random....... here u told amplification should be random...... but in whole video u were talking about the designing of the primers according to the sequence information........ so my question is that... if process is random amplification then what is the need of designing of specific primers according to sequence information??? As per my knowledge in the method of RAPD we use random primers without the prior knowledge of sequence information....... please clarify this....... thank u...
Same question i have asked in the previous video...we don't need the sequence information. For SNP , yes we need sequence info
Ll
This is a confusing process. Why did you sequence the target DNA you extracted from the gel and then still need to do a restriction digest? Surely the sequencing would give you the final answer as to whether or not there is a change, you could just align it?
During the initial part of the video, you said that the 'mutated' sequence will not allow the primer to bind properly, hence we get lighter bands of less concentration. But what if the mutation occurs upstream of the primer binding site? this would mean that the primer will bind properly and also we will also get a complete band because we will have reserves of all four dNTPs anyway. What will be the result then?
my question is that... if process is random amplification then what is the need of designing of specific primers according to sequence information??? As per my knowledge in the method of RAPD we use random primers without the prior knowledge of sequence information....... please clarify this....... thank u...
but in rapd we use the random dna fragments so how the sequences will be added because we don't know the exact sequences?
I mean to say that how the specific primers Wii be added if the DNA sequences are random ?
please tell if template DNA is mistakenly mixture of 2 genotypes but we only want to amplify one genotype... then it'll show mutation ???? or pcr product will not form ????
ek request hai, Sir aap apne poorane videos ko phir se nai videos daaliye sir in sab ya volume kam hoti hai aur user interface thoda dull hai. (Thank u for all your videos)
An interesting clip but I will argue strongly with your differentiation between polymorphism and mutation (from 6:50 to 7:50 min). From a general consensus polymorphism is a result of mutation!
YEAH...its the result of positive mutations
How I can desing primers to RAPD's?
There any software? Thank you.
Excellent video.
Although I really like his videos and his way of explaining the things but in this case the information regarding primer is wrong. Kindly note that RAPD markers are DNA fragments from PCR amplification of random segments of genomic DNA with single primer of arbitrary nucleotide sequence.
I was wondering that too. So the sequence of primer isnt known to us right?
can you please tell how much is the length x width of the white board you use
Nooooo, RAPD doesn't use two primers, it only uses one and it is not a specific one, it's and arbitrary one that binds randomly, that's why it's called RANDOM. The amplification of DNA sequences coming from multiple individuals using the same primer can emphasize the differences after running the gel, the absence of a band in the gel might indicate a SNP in the region that the primer binds, because the primer was unable to bind to that region and thus it couldn't be amplified like in the case of the individual that lacks the SNP in the same region.
Trastorno
You are teaching is very good sir....
You're welcome
Tq sir...
This has been very helpful. Thank you, sir
thank you for such simple and informative video
thank u for nice presentation. i have one some queries 1. why should the length of RAPD markers is 10 base pair and why the length of SSR markers is more than 20 base pair
Scar is also the part of Rapd or not
You are saying here RAPD is dominant marker because it is able to guess or bind with only dominant gene instead of recessive so it's totally wrong....
hello Suman. it is really very informative.bt my question is here you took the example of same gene sequence of two individuals? i.e. one is normal and another is mutated ?
hello. yes he used one normal gene and the mutated gene. rapd is used for the identification of mutations
Dear Sir may God give you a long capable life .....SO YOU CAN GIVE YOUR AS SUCH CONTRIBUTION ..... Thanks
simply incredible way of teaching
+Sp Senthil thank you very much
Are the DNA sequences produced, double stranded or sıngle stranded?
BUT HOW TO DETETERMINE THE SHORT TEMPLATE OF HUMEN 2
thank you! your videos have helped me a lot! Y don't you get a dedicated microphone..
thank you very much sir...... nice explaination, got my doubts clarified....
SNP variation as in told in case of two human being is in coding gene or always in nocoding DNA means these rapid are non coding or coding region
Thank you for this clear lecture!
Thank you soooo much you really helped me good luck
You're welcome
super lecture ,i understood it well,i m a medical student
thanks sir...very helpful video
Thank you very much.
thank you sir ,like ur way of teaching,chanceless ....
+Sp Senthil thank you.
Please Add subtitle to the video. I'm not good at English
I hope you can help me more
I have this project about SNP and microsatellite mainly the detection methods can we say this method in the video is a detection technique as TaqMan Snp and others.
I hope you can help me all the best (3 days to my presentation ) 😢😦
Thank you sir you are doing a good job!!! :)
awesome sir
simple and crispy....
+rajith reddy all the best
is this the same as RACE; rapid amplification of cDNA ends
Sir kindly explain SCAR in a more detailed way...
When people are busy in the things in present time you have been doing this for 7yrs(or may be more). You are more advance than many others.
It's been 10 years on UA-cam
@@shomusbiologyofficial Congratulations to u and ur channel for "a successful decade" in UA-cam...
sir thank u for this video. u explained very well..
Wonderfull...
I would like to see some videos about QTLs too
Regards
was really informative...
The amplification I want to goo🤣.. That's so funny but jokes apart u r genius
Thanks man, very clear
Very helpful...thank you
awesome sir.... i m a student of B.sc agriculture.. n interested in Plant biotechnology...in M.Sc.. could u plz assist me
Thnx a lot sir❤️❤️❤️
You're welcome
very helpful ....thank you soooo very much....
It was useful, thank you .
Thank you!!
great job !!!
Thank you
Thanks very much!
İ was expecting no amlpification for polymorphism.
Thanku sir
thank you sir. as always u r d best
Tq so much
You're welcome
u r great man...keepp it up
Thanks sir
very interesting thanks!
Hindi me bhi btaiye please sir
New Hindi videos are coming
many wrong concepts u discuss like mutation and polymorphism diffrence and about primer , rest of the things r good
Please post the facts slightly elaborated so that other people can find it useful.
No no it is very welp
greattttttttttttttttttttttttt
:) thanks .
Who else thought this was a video of Relative Afferent Pupillary Defect👍
Awesome sir