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We report the design and development of a data acquisition and control system for high-speed acquisition of weak, backscattered differential signals and synchronized sequential operation of all subsystems of a tunable, mid-infrared Differential Absorption Lidar (DIAL) system. The presence of a low-level concentration of chemical species results in weak differential return signals. The differential signal also varies dynamically with respect to background atmospheric conditions. The challenge is to measure this low level of differential signal with high resolution and also control the sequence of operation of subsystems such as lasers and scanners for real time testing and evaluation in open field conditions. The concentration spread of the chemical species varies rapidly with distance. In order to capture this spatial variation, the lidar signal should be sampled and digitized at a high sampling rate. A customized Peripheral Component Interconnect based data acquisition of a 12-bit resolution, 30 Mega samples per second sampling rate, and an industrial personal computer-based control system has been realized. Detection algorithms and the firing sequence of the laser have been developed indigenously and implemented in the LabVIEW platform. The developed graphical user interface has various modes of operation as per user requirement and is capable of executing automatic operations for the developed DIAL system in order to detect and identify the chemicals. The performance has been evaluated by detecting the chemical species thiodiglycol at 800 m using 3190 and 3300 nm (online and offline wavelengths) with a differential cross-section of 2.5 × 10−23 m2. Similarly, methane has been detected and quantified with concentration of 2.2 ppm up to 300 m using 3316 and 3326 nm (online and offline wavelengths, respectively).