![]() ![]() To ensure that the correctly fused product was obtained (4 th lane), the resulting fusion was PCR purified and directly microinjected in the germline of wildtype (N2) animals and the resulting expression pattern examined from stable transgenic animals. Using genomic DNA as template, the promoter region of myo-3 was amplified (2 nd lane), and fused to the GFP coding region containing the unc-54 3′UTR which was amplified from the plasmid pPD95.75 (3 rd lane). (A) Primers were designed using STITCHER 2.0 to fuse the promoter region of the C. STITCHER 2.0 is freely available to all users without signup or login requirements and can be accessed at the following webpage: PubMed Disclaimer 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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