Flipped instruction using online enrichment is a popular way to enhance active learning in the laboratory setting. Graduate student teaching assistants at University of California, Irvine flipped an upper division undergraduate neurobiology and behavior lab using the new online software platform "Rocketmix." The following research study compares the impact of pre-lab online instruction (front flipping) and post-lab online instruction (back flipping) on student exam performance. We describe a novel method for unbiased categorization of exam questions by degree of difficulty. Multi-choice instruction encourages students to consider all distractors and discourages verbal cues and process of elimination techniques. Eighteen identical questions were evenly distributed across exam versions with multiple choice instruction (single answer) or a more challenging multi-choice instruction (more than one answer). Student performance on multiple choice questions were used to categorize the degree of difficulty of questions that were presented in multi-choice format. Our findings reveal that pre-lab instruction resulted in better student performance compared with post-lab instruction on questions of moderate difficulty. This effect was significant for both male and female students. Student survey data on the flipped lab format is provided, indicating that students appreciated the online instructional modules, finding them both informative and useful during lab exercises and exams.