background: th e second leading cause of cancer deaths in women is breast cancer. germline mutations in susceptibility breast cancer gene brca1 increase the lifetime risk of breast cancer. eighty-one large genomic rearrangements (lgrs) have been reported up to date in brca1 gene, and evaluation of these rearrangements helps with precise risk assessment in high-risk individuals. in this study, w...

NASBA is an isothermal nucleic acid amplifi cation method that is particularly suited to detection and quantifi cation of genomic, ribosomal or messenger RNA. The product of NASBA is single-stranded RNA of opposite sense to the original target. The fi rst developed NASBA methods relied on liquid or gel-based probe-hybridisation for post-amplifi cation detection of products. More recently, real-...

The development of instruments that allowed real-time monitoring of fl uorescence within PCR reaction vessels was a very signifi cant advance. The technology is very flexible and many alternative instruments and fl uorescent probe systems have been developed and are currently available. Real-time PCR assays can be completed very rapidly since no manipulations are required post-amplifi cation. I...

We present a protocol for effi cient amplifi cation of a large number of DNA targets using a single-tube polymerase chain reaction (PCR) based on PCR suppression. This method allows amplifi cation of each DNA target with only one target-specifi c primer whereas the other primer is common for all targets. Thus, this approach requires n+1 primers for n targets instead of 2n in conventional PCR an...

Mercury is one of the more toxic elements to living organisms. It is encountered at relatively low concentrations in the environment but with its amplifi cation through the food chain, the fi nal concentration in some foods can be relatively high. In addition, its toxicity is not only linked to its total concentration but is also dependent on the species. Mercury speciation in food analysis is ...

Peptide nucleic acid (PNA) oligomers can be employed as site-specifi c openers of the DNA double helix to locally expose a designated marker sequence inside duplex DNA. The opened DNA site is then hybridized to a circularizable oligonucleotide probe, which is subsequently closed by DNA ligase. This way, the marker sequence from the DNA duplex of interest can be isothermally amplifi ed by a vari...

and second-round PCR were 94°C for 3 min, followed by 40 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 2 min. Expected amplifi cation products were 458 bp (PCR-1) and 304 bp (PCR-2). Using dilutions of a synthetic template corresponding to the target sequence, we estimated the sensitivity of the amplifi cation assay to be <5 copies of target sequence by limiting-dilution assay. Negative ...