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| PCR - Polymerase Chain Reaction Forum PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory. |
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#1
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| hi, im doin RACE, which now im facing some problems. does anyone knows why GSP1 and GSP2 doesnt anneal to the adapter. could anyone give me opinion on why its happen and to overcome this problems? TQ in advance Regrads, Q |
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#2
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| I'm also working on RACE, and had some samples successfully amplified. I hope it's a small typo mistake you made in your post, if not then I would suggest you go through the manual carefully again on how RACE is supposed to work; 1. GSP (gene-specific primer) is the primer you design to complement to a region of your desired gene. You use GSP together with a supplied 5' or 3'-primer in the kit to carry out your 5' or 3'-RACE. Your GSP is NOT supposed to anneal to the adapter; the primers that come with the kit is supposed to do that. 2. RACE is definitely much more trickier than PCR. If you don't see any bands or smears, don't give up yet. Use the RACE products as template for a subsequent nested/internal RACE. If you're lucky, you'll be able to see some bands or smears. |
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