Optimization of DNA extraction from low-yield and degraded samples using the BioRobot EZ1 and BioRobot M48.
Robotic extraction of DNA from dilutions of blood and semen using either the BioRobots EZ1 or BioRobots M48 consistently produced lower recoveries than standard organic extractions of the same samples. In an effort to increase the efficiency of robotically extracted DNA, glycogen and carrier RNA were added following cell lysis. The addition of glycogen, postlysis, resulted in no improvement in DNA recovery with the BioRobot EZ1. However, when carrier RNA was added to the cell lysate of limited and degraded samples extracted on the EZ1 or the M48, DNA recoveries dramatically increased four- to 20-fold. DNA yields obtained by robotic extraction in the presence of carrier RNA were as high, or higher, as those obtained by organic extraction lacking carrier RNA, while experiments that utilized carrier RNA in both types of extractions showed increased sensitivity for both methods. Furthermore, carrier RNA substantially increased the recovery of fragmented DNA with the EZ1.
Kishore
Ram
R
Jan Bashinski DNA Laboratory, California Department of Justice, 1001W. Cutting Blvd., Suite 110, Richmond, CA 94804, USA.
Reef Hardy
W
W
Anderson
Vince J
VJ
Sanchez
Nick A
NA
Buoncristiani
Martin R
MR
D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D005554:Forensic Medicine / Q000379:methods; D006003:Glycogen; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D012343:RNA, Transfer; D012371:Robotics; D012661:Semen / Q000737:chemistry; D020080:Tandem Repeat Sequences
D006003:Glycogen; D004247:DNA; D012343:RNA, Transfer