2 replies to this topic

[chat25]

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

[fishdoc]

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.

[chat25]

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......