Targeted rapid amplification of cDNA ends (T-RACE)—an improved RACE reaction through degradation of non-target sequences

Amplification of the 5′ ends of cDNA, although simple in theory, can often be difficult to achieve. We describe a novel method for the specific amplification of cDNA ends. An oligo-dT adapter incorporating a dUTP-containing PCR primer primes first-strand cDNA synthesis incorporating dUTP. Using the Cap finder approach, another distinct dUTP containing adapter is added to the 3′ end of the newly synthesized cDNA. Second-strand synthesis incorporating dUTP is achieved by PCR, using dUTP-containing primers complimentary to the adapter sequences incorporated in the cDNA ends. The double-stranded cDNA-containing dUTP serves as a universal template for the specific amplification of the 3′ or 5′ end of any gene. To amplify the ends of cDNA, asymmetric PCR is performed using a single gene-specific primer and standard dNTPs. The asymmetric PCR product is purified and non-target transcripts containing dUTP degraded by Uracil DNA glycosylase, leaving only those transcripts produced during the asymmetric PCR. Subsequent PCR using a nested gene-specific primer and the 3′ or 5′ T-RACE primer results in specific amplification of cDNA ends. This method can be used to specifically amplify the 3′ and 5′ ends of numerous cDNAs from a single cDNA synthesis reaction

Thermal performance of different working fluids in a dual diameter circular heat pipe

In this paper, heat transfer performance of a 40 cm-length circular heat pipe with screen mesh wick is experimentally investigated. This heat pipe is made of copper with two diameters; larger in the evaporator and smaller in the adiabatic and condenser. Three different liquids including water, methanol, and ethanol are separately filled within the heat pipe. Low heat fluxes are applied (up to 2500 W/m2) in the evaporator and constant temperature water bath is used at three levels including 15, 25, and 35 °C in the condenser. Results demonstrate that higher heat transfer coefficients are obtained for water and ethanol in comparison with methanol. Furthermore, increasing heat flux increases the evaporator heat transfer coefficient. For the case of methanol, some degradation in heat transfer coefficient is occurred at high heat fluxes which can be due to the surface dryout effect. Increasing the inclination angle decreases the heat pipe thermal resistance