![]() Unlike STITCHER, STITCHER 2.0 allows the users to customize more features, and provides more robust control regarding overlap complementarity. Here we present a significant upgrade called STITCHER 2.0, which considers more algorithmic parameters during primer design, and provides numerous visual guides and output files that provide a better resolution of the designed primers and starting sequence for the end-user ( Fig. ![]() We have previously developed a web-based tool called STITCHER that automates the design of primers for many overlapping PCR applications 5. Some of the applications of overlapping PCR include gene synthesis by assembly PCR, reporter gene fusion using GFP or RFP based fluorophores, and detection experiments through techniques such as loop-mediated isothermal amplification (LAMP) 2, 3, 4. STITCHER 2.0 is freely available to all users without signup or login requirements and can be accessed at the following webpage: Overlapping PCR is a common technique used by molecular biologists in various fields that allows the user to ‘stitch’ together various individual DNA fragments 1. These result files provide greater control and insight during experimental design and troubleshooting. Unlike STITCHER, STITCHER 2.0 considers diverse algorithmic parameters, and returns multiple result files that include a facility for the user to draw their own primers as well as comprehensive visual guides to the user’s input, output, and designed primers. STITCHER 2.0 is a newly designed web tool that automates the design of primers for overlapping PCR. ![]() Here we present STITCHER 2.0, which represents a substantial update to STITCHER. Previously, we have reported a web based tool called STITCHER that provides a platform for researchers to automate the design of primers for overlapping PCR applications. Overlapping polymerase chain reaction (PCR) is a common technique used by researchers in very diverse fields that enables the user to ‘stitch’ individual pieces of DNA together.
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