The reaction continues until accumulation of stem-loop DNA structures with several inverted repeats

of the target and cauliflower-like structures with Entospletinib in vivo multiple loops formed by CHIR98014 mouse annealing between alternately inverted repeats of the target in the same strand. The reaction takes less than an hour, and produces 109 copies of the target DNA [21]. Due to its significant advantages over PCR, since its development, LAMP has been found in many applications in the molecular diagnosis including detection of pathogens, genetically modified organisms, embryo sex detection, and tumor detection [22]. One advantage is that in LAMP method, a particular bacterial DNA polymerase, named Bst DNA polymerase, is used for amplification of the target in isothermal condition without application of any specific thermal cycler. Because this enzyme denatures simultaneously the substrate double-strand DNA and subsequently synthesizes

products, there is Adriamycin molecular weight no need for pre-denaturation of target dsDNA, especially by heat, which is used commonly in PCR technique. The amplification thus can be performed in a simple water bath or other heating tool in isothermal condition. Secondly, in spite of PCR, products of LAMP can be detected visually by the naked eye through turbidity or adding DNA intercalating dyes (e.g., SYBR green) without any need for specified equipment (gel electrophoresis and UV gel documentation systems which are necessary in PCR). Also, since the amplification and detection are performed in the same tube, unlike PCR, LAMP can be performed in closed reaction tubes and it minimizes any cross-contamination risk while using multiple samples [21, 23–25]. Besides this advantage, LAMP is not as sensitive as PCR toward inhibitors [26]. It thus contributes to lowering of LAMP costs compared with PCR. The third advantage is the higher specificity why and sensitivity of LAMP over PCR. Unlike common PCR, performed using one pair of primer, LAMP requires at least two pairs of primer and, thus, increases the chance of specific

recognition and amplification of target DNA compared to PCR. Higher sensitivity of LAMP originates from the unique mechanism of amplification, known as loop-mediated amplification, which produces huge amplicons consisting of repeated loop-shaped and dumbbell-like DNA structures. This type of amplicons allows easier detection of positive results and lowers the limit for detection. Therefore, it increases sensitivity of LAMP in comparison with PCR. The fourth advantage of LAMP is its speed over PCR. The overall time for carrying out LAMP is about 1 h compared with 2 to 4 h in PCR [21, 23–25] Moreover, the rate of LAMP reaction can be increased by using two additional primers, called loop primers [27]. Therefore, LAMP is considered a more rapid molecular technique compared with PCR. LAMP can be performed in a high-throughput format for simultaneous analysis of large-number samples in 96-well microplate [28].