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reconstitution of primers - (Sep/03/2009 )

Hi all,

I need to know how to reconstitute a primer. actually i'm a fresher to this feild and this is the first time i'm running a PCR. the primer we bought has given the details as,

forward - Tm 64 oc 82.9 nmol
reverse - Tm 65.9 oc 63.4 nmol

i want to know how to use this primer to perform Gradient PCR.

Please help me out on this...

thanks

-chat25-

chat25 on Sep 3 2009, 11:59 AM said:

Hi all,

I need to know how to reconstitute a primer. actually i'm a fresher to this feild and this is the first time i'm running a PCR. the primer we bought has given the details as,

forward - Tm 64 oc 82.9 nmol
reverse - Tm 65.9 oc 63.4 nmol

i want to know how to use this primer to perform Gradient PCR.

Please help me out on this...

thanks


Resuspend in water or tris to a final concentration of 100 uM. Use this as a stock solution and make a working solution of 10 uM, then add 2.5 ul of that to a 50 ul PCR reaction to get a final concentration of 0.5 uM per primer, or adjust to whatever concentration you want.

-fishdoc-

fishdoc on Sep 3 2009, 10:07 AM said:

chat25 on Sep 3 2009, 11:59 AM said:

Hi all,

I need to know how to reconstitute a primer. actually i'm a fresher to this feild and this is the first time i'm running a PCR. the primer we bought has given the details as,

forward - Tm 64 oc 82.9 nmol
reverse - Tm 65.9 oc 63.4 nmol

i want to know how to use this primer to perform Gradient PCR.

Please help me out on this...

thanks


Resuspend in water or tris to a final concentration of 100 uM. Use this as a stock solution and make a working solution of 10 uM, then add 2.5 ul of that to a 50 ul PCR reaction to get a final concentration of 0.5 uM per primer, or adjust to whatever concentration you want.



thanks alot......

-chat25-