Realtime PCR

[rangermat]

I am really new to qpcr and I need to set up this pcr for 2 different DNA fragments with concentration of 0.1nM. Total volume of each fragment is 5ul.Was told to make a diltution series between 10pM to 0.1nM. I am really confused as to what to do. can someone please help or advice?

[Phototoxin]

Essentially you need to amplify the DNA at different concentrations.

you use 5µl of the end solution I'm guessing.
You would seeminglu need to make serial dilutions.


Nanomolar nM 10^-9 molar 0.000,000,00X
Picomolar pM 10^-12 molar 0.000,000,000,00x (commas to help counting dont actually put them in!)

10 pM =0.000,000,000,010 M
0.1 nM=0.000,000,000,1 M

essentially convert the nanoMoles into picoMoles

0.1nM = 100 pM
10pM = 10pM ;-)

So you need a dilution series of 10 and 100pM. I would need to know what you started with to give exact examples but if you had 100pM/0.1nM solution and you need to make 10pM solution you'd do a 1 in ten. Get 45 units of buffer and add 5 units of your 100pM solution. This would dilute it by a factor of 10. (5+45=50, 5 is 10% of 50)

The units depend on what you're working with... ml, µl or whatever.

Hope that helps.

[rangermat]

Thanks for your input...was able to figure it out later! Effort really appreciated!