Abstract: Methods and kits for use in the practice of reverse transcriptase and for one-tube practice of reverse transcriptase and polymerase chain reaction are described. Use of an RNA template having a known conserved area of at least about 40 base pairs, a reverse transcription primer suitable for preparing DNA from the conserved area of said RNA template, and a reverse transcriptase having an elongation rate of at least 40 base pairs per second and a processivity of at least 75 base pairs allows for rapid reverse transcription of the conserved area. Use of PCR primers having 3? ends which are separated by about 1 to about 120 base pairs, DNA polymerase having an elongation rate of at least 60 base pairs per second and a processivity of at least 150 base pairs together with PCR reaction components allows for rapid amplification of DNA corresponding to the conserved area of the RNA.

Abstract: Methods and apparatus for effecting rapid thermocycling in connection with the polymerase chain reaction are disclosed. A sample assembly having a relatively small thermal mass is heated to desired PCR operating temperatures, and a separate cooling assembly is used to rapidly lower the temperature as required. In one embodiment, a sample assembly with an integrated heating element is isolated from a relatively large thermal mass cold sink when the temperature of a sample is to be raised or maintained, and brought in contact with the cold sink when the temperature is desired to be lowered.

Abstract: Apparatus and methods for exciting and detecting fluorescence in samples are disclosed. In one embodiment, a sample holder for holding a plurality of samples is provided together with an optical manifold having an excitation source, a photo receiver, or both, for each of the plurality of samples. In another embodiment, the optical manifold contains only the excitation source or a photo receiver, and the other is associated with the sample holder. This system permits for rapid excitation and measurement of fluorescence without the use of moving parts and without any opto-mechanical or electronic disturbance. It exhibits an exceptional signal to noise ratio, which permits it to differentiate between very low level differences in fluorescence.