I am a bit confused.
I did RNA extraction and the yield is 17.5 ng/ul (its low). My final volume is 30 ul
Next i did a DNAse treatment.
Now, i want to do a cDNA synthesis followed by a qPCR.
For cDNA synthesis i am using Thermo scientific cDNA synthesis kit and for qPCR i am using Maxima SYBR green kit.
For the cDNA synthesis the kit says use 1 pg - 5 microgram (ug) of template RNA. I want to know how can i convert 17.5 ng/ul to 1 pg or 5 ug.
Next, for the qPCR master mix, it says use < or = 500 ng of template DNA. So, how much cDNA should i use.
Please let me know how do i make these calculations.