Polymerase resistance to polymerase chain reaction inhibitors in bone*.
نویسندگان
چکیده
Amplification of DNA from aged or degraded skeletal remains can be a challenging task, in part due to naturally occurring inhibitors of the polymerase chain reaction. PCR inhibitors may act by inactivating a polymerase itself, or compete with or bind other reaction components, although various polymerases may be differentially susceptible to such insult. In this study, ten thermostable polymerases from six bacterial species were examined for their ability to amplify DNA in the presence of bone-derived or individual PCR inhibitors. Two polymerases, one from Thermus aquaticus and one from Thermus thermophilus, showed lower susceptibility to inhibition from bone, while polymerases from Thermus flavus were highly susceptible. Addition of bovine serum albumin improved the activity of most of the enzymes. Taken together, the results indicate that thermostable DNA polymerases have different susceptibility to bone-derived PCR inhibitors, and that those most often used in forensic laboratories may not be optimal when working with DNA from skeletal remains.
منابع مشابه
Detection of tetracycline resistance genes in bacteria isolated from fish farms using polymerase chain reaction
Five common tetracycline resistance genes tet(A), tet(B), tet(M), tet(O) and tet(S) were studied by polymerase chain reaction in 100 bacteria isolated from Iranian fish farms. In the antibiogram test most of the bacteria were either intermediately or completely resistant to tetracycline. Nine isolates out of 46 Aeromonas spp. contained eithe...
متن کاملTyping of HLA Class I by Polymerase Chain Reaction-Sequence Specific Oligonucleotide Primer (PCR-SSOP) Technique in Iranian Cord Blood Donors
Background: HLA compatibility between transplant donor and recipient is one of the major determinants of transplant outcome. Objective: To determine HLA class I by PCR- Sequence-Specific Oligonucleotide Probe (PCR-SSOP) in cord blood donors. Methods: Genomic DNA of 142 cord blood samples registered at the Cord Blood Bank of Iran at Hematology, Oncology, and Bone Marrow Transplantation Researc...
متن کاملDetection Of Toxoplasma Gondii and Human Cytomegalovirus DNA in Blood from Transplant Recipients Using Multiplex Nested Polymerase Chain Reaction
Evidences from many studies suggested a polymerase chain reaction (PCR) as a valuable method for diagnosing infectious disease in the transplant recipients. We used this method for detection of Toxoplasma, gondii and human cytomegalovirus in blood specimens from patients after bone marrow or kidney transplantation. DNA of both infectious agents were detected using two separate sets of nested pr...
متن کاملDetection of Nocardia Asteroides Complex in Clinical Isolates by Real-Time Polymerase Chain Reaction
Background and Aims: Nocardia asteroides complex is the most common cause of infectious diseases due to nocardiosis. Interspecies differentiation of Nocardia genera is essential for prognosis and timely proper treatment, as well as for epidemiological studies. Since each genus has its own antibiotic resistance, precise careful diagnosis is of prime importance. As compared to biochemical and phe...
متن کاملGenetic properties of blaCTX-M and blaPER β-lactamase genes in clinical isolates of Enterobacteriaceae by polymerase chain reaction
Objective(s):blaCTX-Mand blaPER are two genes that encode class A extended-spectrum β-lactamases (ESBLs) and can be responsible for therapeutic problems. This study was carried out to evaluate the molecular properties of these genes in clinical isolates of Enterobacteriaceae by polymerase chain reaction (PCR), restriction digestion and sequencing. Materials and Methods: During six months, start...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of forensic sciences
دوره 54 5 شماره
صفحات -
تاریخ انتشار 2009