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multiplex PCR for 9 exons


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5 replies to this topic

#1 lula

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Posted 25 March 2005 - 01:55 AM

hi

im working a multiplex PCR o specify deleions in nine exons ,,,,,
i usually make the reacion on 3 tubes ,,,each with 6 primers
im getting sharp bands but tthe trouble is that only one of the nine bands (he biggest size )is thin

does this count for a band ??????? even though its thin ???????? it its appearing in its right position but just very thin

#2 Scott

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Posted 29 March 2005 - 12:18 AM

Hi lulu,

Two ways to tell if what you are seeing is real. The first is to clone it and sequence it. The second is a control reaction where you leave the specific primers that would give that band out of the reaction, if the band disappears than you know it is due to that primer pair and is more than likely real.

Cheers,

Scott

#3 lula

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Posted 29 March 2005 - 08:12 AM

hi scott
thanks for ur reply,,,
i dont have a sequencer but the control method u mentioned seems god enough and applicable

i did the same PCR reaction on another case and the same band appeared and again it was thin
but i well try ur method its more precise
thanks

Edited by lula, 29 March 2005 - 08:14 AM.


#4 lula

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Posted 30 March 2005 - 10:52 AM

hi scott
i used the primer method u told me abut and it worked great :)
now i can give the result to the pateient with a clear happy conciuos
thanks alot

#5 lula

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Posted 30 March 2005 - 10:53 AM

hi scott
i used the primer method u told me abut and it worked great :)

thanks alot

#6 Scott

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Posted 31 March 2005 - 01:30 AM

Glad I could help

:)




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