Step-by-Step Method Details Your Major Step One Timing: [s, min, h, days, or weeks] [Include a brief description about what this major step accomplishes; this will help other researchers repeat and troubleshoot the protocol.] [First step of major step one. Use numbers to order steps.] [Sub-steps under the first step. Use letters for ordering sub-steps. Align the sub-steps slightly to the right to be better differentiated from steps.] [Sub-step two, as needed.] [Third-level sub-steps under second-level sub-steps. Use Roman numerals for ordering. Align the third-level sub-steps slightly to the right to be better differentiated from second-level sub-steps.] [Third-level sub-step two, where needed.] [Second step of major step one] Critical: [Particularly important instructions or factors should be called out in this format. Please provide figures or videos for guiding critical steps. Click here to see examples of useful figures.] Your Major Step Two Timing: [s, min, h, days, or weeks] [Include a brief description about what this major step accomplishes. This will help other researchers repeat and troubleshoot the protocol.] [First step of major step two. Use numbers to order steps.] [Sub-steps under first step. Use letters for ordering sub-steps. Align the sub-steps slightly to the right to be better differentiated from steps.] [Sub-step two, as needed.] [Second step of major step two] Troubleshooting 1 Optional: [Call out optional steps in this format; optional steps are not numbered.] Note: [Please include general notes or explanations that would be helpful in this format.] Pause Point: [If relevant, list potential pause points with time intervals in this format.] Use this format to present PCR reactions (if relevant).