Use of AmpliWax to optimize amplicon sterilization by isopsoralen.

Journal of Clinical Microbiology
M De la ViudaJ Aslanzadeh

Abstract

The photochemical inactivation of amplicons by isopsoralen (IP-10) has been suggested as a possible means to prevent PCR carryover contamination. To evaluate the technique, serial dilutions of amplicons (10(11) to 10(3)) from the Borrelia burgdorferi OSP A gene were amplified in the presence of 0, 25, 50, and 100 micrograms of IP-10 per ml for 45 cycles. The PCR products were exposed to UV light for 15 min to activate IP-10 and sterilize the amplicons. One microliter of each sterilized sample was reamplified for an additional 45 cycles. The PCR products were then resolved in an agarose gel, blotted onto a nylon membrane, and probed with an alkaline phosphatase-conjugated chemiluminescent probe. Although IP-10 at concentrations of 50 and 100 micrograms/ml effectively sterilized up to 10(11) amplicons, the compound was inhibitory to PCR. IP-10 at a concentration of 25 micrograms/ml had slight inhibitory effect on PCR and did not completely sterilized all of the amplicons. Therefore, in subsequent experiments AmpliWax was substituted for mineral oil, and PCR was performed on 10(9) to 10(3) amplicons as described above. Following the amplification, the PCR tubes were cooled to solidify the AmpliWax and inoculated with various conce...Continue Reading

References

Oct 29, 1992·The New England Journal of Medicine·L S Tompkins
Sep 1, 1992·International Journal of Food Microbiology·L RossenO F Rasmussen
Feb 1, 1992·Molecular and Cellular Probes·D E Dwyer, N Saksena
Aug 1, 1991·Journal of Virological Methods·J C FoxV C Emery
Jan 11, 1991·Nucleic Acids Research·S T IsaacsG D Cimino
May 11, 1991·Nucleic Acids Research·Y S ZhuJ E Hearst
Jan 4, 1990·Nature·G Sarkar, S S Sommer
Jun 28, 1990·Nature·G D CiminoY S Zhu
May 18, 1989·Nature·S Kwok, R Higuchi
Mar 1, 1994·Journal of Clinical Microbiology·L F KoxA H Kolk
Apr 1, 1993·Molecular and Cellular Probes·J Aslanzadeh

❮ Previous
Next ❯

Citations

Dec 17, 1998·Journal of Clinical Microbiology·G A FahleS H Fischer

❮ Previous
Next ❯

Related Concepts

Related Feeds

ApoE, Lipids & Cholesterol

Serum cholesterol, triglycerides, apolipoprotein B (APOB)-containing lipoproteins (very low-density lipoprotein (VLDL), immediate-density lipoprotein (IDL), and low-density lipoprotein (LDL), lipoprotein A (LPA)) and the total cholesterol/high-density lipoprotein (HDL) cholesterol ratio are all connected in diseases. Here is the latest research.