background: splicing by overlap extension (soe) pcr is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function.objectives: we introduced a nested-soe-pcr (n –soe-pcr) in order to increase the specificity and generating mutations in a gene by soe-pcr.  materials and methods: genomic dna from bacillus thermoca...

The contamination of apheresis products with tumor cells was evaluated in patients undergoing autologous peripheral blood stem cell transplantation for germ cell tumors. A blinded, retrospective analysis was performed on 63 apheresis products from 28 patients using the PCR and primers for beta human chorionic gonadotropin (beta-HCG). Of the 20 patients with beta-HCG-secreting tumors, 8 apheresi...

By incorporating dUMP residues into the 5' end of PCR primers, one can generate products which, after treatment with uracil DNA glycosylase (UDG), contain 3' overhangs. These overhangs can be annealed to vector molecules with complementary overhangs generated in a similar fashion and transformed directly into Escherichia coli without the need for ligase. We have tested this method of ligation-i...

An automated system for polymerase chain reaction (PCR) amplification and detection combats false-positive results caused by "PCR product carryover." The system uses a single vessel for both PCR amplification and the subsequent detection of PCR products, eliminating the need to handle PCR products in an open environment and risk product carryover. The sample and PCR reagents are introduced into...

A polymerase chain reaction-gold magnetic nanoparticles lateral flow assay (PCR-GoldMag LFA) has been developed via integrating multiplex amplification refractory mutation system PCR (multi‑ARMS‑PCR) with GoldMag‑based LFA for the visual detection of single‑nucleotide polymorphisms (SNPs). This assay was applied to genotype Apolipoprotein E (ApoE). ApoE genotyping is important due to the predic...

The efficacy of six different sets of primers targeted against 16S rRNA and virulence genes such as 'iap', 'hly' and 'prf' was evaluated in separate PCR assays. The primer pairs targeted against 16S rRNA resulted into amplification of 1.2 kb PCR product. However, sets of primers targeted against different regions of 'iap' produced 371 and 660 bp PCR products, respectively. The primer pair targe...

In order to determine whether mixed infections of human herpesvirus 6B (HHV-6B) occur in immunocompetent and immunocompromised individuals, we examined the copy numbers of telomeric repeat sequences (TRS) of clinical isolates. In clinical isolates obtained from patients with exanthem subitum caused by primary HHV-6B infection, PCR products with HHV-6B TRS ranging between 400 and 800 bp were amp...

Herpes B virus DNA was specifically amplified by PCR, targeting the regions that did not cross-react with herpes simplex virus (HSV). The amplified products, which were shown to be highly genetic polymorphisms among herpes B virus isolates, were identified by microplate hybridization with probes generated by PCR. The products immobilized in microplate wells were hybridized with the biotin-label...

Rapid pathogen testing is vital to the food industry. Enzyme immunoassays (EIA) provide reliable negative results in 48 h, but a presumptive positive (suspect) EIA result must be confirmed by traditional culture methods, requiring an additional 72 h. Polymerase chain reaction (PCR) testing technology is accepted as an accurate diagnostic tool. However, traditional PCR techniques can require sev...

The recently developed Inter-Simple Sequence Repeat PCR (ISSR-PCR) or microsatellite primed PCR or Simple Sequence Repeat (SSR)-Anchored PCR technique detects polymorphic markers in a wide variety of genomes. Usually the ISSR primers are either 5' end-labeled with gamma[32P]ATP or one of the alpha[32P] labeled dNTPs is added to the PCR reaction and the PCR products are resolved on PAGE and auto...