This research aims to produce valid and reliable prior knowledge questions on the concept of cellular respiration online by utilizing Google form media. The questions developed in this study are multiple choice questions with a four-tier test type. The sample in this study were students who took general biology courses at Faculty of Teacher Training and Education (FKIP) of Universitas Sriwijaya. Samples play a role in the evaluation process after prototype I was produced, at one-to-one (n = 13) and small group (n = 23) stages. The instrument used in the data collection process was a walkthrough, question readability questionnaire and questions. The questions given to the sample consisted of multiple choice questions on the grounds of open-ended questions (the development stage) and four-tiered multiple-choice test questions (the small-group evaluation stage). The use of the Goggle form in this study was carried out at the stage of giving open-ended question questions and the evaluation process at the one-to-one and small group stages. The results of this study indicate that the prior knowledge questions that have been developed are categorized as valid with an expert review rating of 80.5% and the reliability of the categorized questions is weak with a value of 0.37. The results of the expert review assessment revealed that the items developed were in accordance with the understanding of the concepts to be measured. Findings from the results of this study can be used as a reference for developing prior knowledge questions with the type of four-tier test to detect students’ understanding before learning is done. Thus the teacher is expected to choose the right learning strategy before the learning process.