I obtained less than 0.5 ng/ul of capture libraries while the amount of individual libraries were as expected; what are the possible issues?
Usually with most of our panels the amount of DNA we quantify after the capture is between 10 and 30 ng/µl. Low yields may be related to several problems:
- incomplete denaturation step
- a decrease of temperature after denaturation and prior to the addition of the probes
- omission of probe addition
- Loss of material during the wash steps (?)
- Problems with post-capture amplification
- Loss of material during cleanup