Need help with Sequencing troubleshooting!

[r.rosati]

I've never seen a 70% isopropanol rinse.

I agree, my brain just processed "70% ethanol" there.

And I also agree that drying on speedvac might be troublesome.

[mchlbrmn]

Thanks Relaxin. I also noticed that the protocol above used more than one volume of isopropanol, while most protocols use 0.7-1.0 volumes, with 0.8x being the ratio I learned.
(70% x72ul) isopropanol / (12ul PCR +30% of 72ul) water

[r.rosati]

Isopropanol volume is ok, I use 80 ul of 75% iPrOH for 20 ul reactions (80*75/100 = 60% final) and that's the same of using 72 ul of 70% iPrOH for 12 ul reaction (70*72/84 = 60% final). This is a correct concentration.