The invention discloses a microfluidic chip fixture and a droplet preparation system. The microfluidic chip fixture comprises a bottom board and a cover board, a containing groove for containing the microfluidic chip is formed in the bottom board concavely, the cover board can detachably cover the bottom board and is at least provided with a first sample storing hole, a second sample storing hole, a reflux and sample-collection hole, a first jointing hole and a second jointing hole, the first sample storing hole is used for corresponding to a first sample injection port, the second sample storing hole is used for corresponding to a second sample injection port of the microfluidic chip, the reflux and sample-collection hole is used for corresponding to a sample outlet of the microfluidic chip, the first jointing hole is used for containing a first joint connected with a gas source, and the second jointing hole is used for containing a second joint connected with the gas source. The microfluidic chip fixture has the advantages that trace quantified sample injection can be directly performed on the cover board, and a traditional external sample injection tube is omitted, so that loss of biological samples in the external sample injection tube is avoided; cleaning and tube replacement are avoided, consumption of the external sample injection tube is reduced and lower cost is achieved.