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Cloning at 6 kb gene


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3 replies to this topic

#1 praveen921

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Posted 07 March 2013 - 05:25 AM

Hello all,
Im trying to Clone a 6KB gene from flower cDNA.Any suggestions before i actually start the work?Im not sure if its gonna be easy amplyfing a 6kn fragment.Is there a good solution to clone this?
Thanks!

#2 mdfenko

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Posted 07 March 2013 - 05:49 AM

use a high fidelity proofreading polymerase in your pcr

Edited by mdfenko, 07 March 2013 - 05:49 AM.

talent does what it can
genius does what it must
i do what i get paid to do

#3 praveen921

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Posted 08 March 2013 - 06:56 AM

Hi i used two polymerases in separate reactions to amplify the 6KB gene
1.Pfu Ultra II DNA Polymerase

2.PrimeSTAR HS DNA Polymerase

I followed the PCR conditions they mentioned in the protocol,but i wasnt able to amplify the gene.What should i do now?
thanks!

#4 mdfenko

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Posted 08 March 2013 - 12:20 PM

check that your primers are appropriate.
talent does what it can
genius does what it must
i do what i get paid to do




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