Premium
Optimization of DNA Extraction from Low‐Yield and Degraded Samples Using the BioRobot ® EZ1 and BioRobot ® M48
Author(s) -
Kishore Ram,
Reef Hardy W.,
Anderson Vince J.,
Sanchez Nick A.,
Buoncristiani Martin R.
Publication year - 2006
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2006.00204.x
Subject(s) - lysis , rna , dna , extraction (chemistry) , microbiology and biotechnology , dna extraction , rna extraction , chromatography , biology , biochemistry , chemistry , polymerase chain reaction , gene
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.