I made an experiment through which i was able to label human genome using two different fluroscent non dye terminators dNTPs , however i need to check the fidelty of the method through a sequncer that can identify my dyes position and then to dectate the compleentary strand of it , as we know the base pairing rule govern nucleotide insertion .
Dear Awadelkareem :
What do your " non dye terminators dNTPs look like?
"different fluorescent non dye terminators dNTPs " and " identify my dyes position "
Do you have dye on your terminator or else?
Thank you!
Rhodamine 12-dCTP and amionallyl dUTP-XX-AF488 . Both are not dye terminator and incorporated in the DNA
They have dyes (Rhodamine 12 and AF488 both are dye).
Do you have any group on the 3'-OH? or they are ddNTPs, not dNTPs.
Wish the best!
Yes Dr.Guobing. they possess OH at the 3 position on the ribose
Sir, if they have free 3'-OH, they are not terminators in general, except the virtual terminators of Helicos' single molecular real time sequencing technology, those virtual terminators have inhibitors linked to the ap dNTPs.
Nature Methods, 2009, 6(8), 593
Good luck!
Dear Mr.Xiang. there is any genetic analyzer that can sequence my tagged DNA !
We made it possible in our noval eperiment to incorporate the above two dyes into homopolymer DNA molecule that reached a length of 3.3GB. our problem is to find a new technology that can sequence pretagged DNA without library prepration or PCR amplification as will as sequencing by synthesis , something similar to nanopore tachnology that read DNA immediately but only using laser .
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