When designing probes for existing primer pairs with NCBI Primer-BLAST software it always cannot consider system with probes, i.e. cannot design the probe (pick internal oligo). This problem also occurs when we try to check our systems with good probe (we can check by primer3 and blast). At the same time, if we tape probe sequence instead of forward primer PrimerBlast does check the system and moreover usually considered it OK.
Does anybody know what additional settings must be done to solve this problem (design probe to pre-existing primer pairs) in PrimerBlast?
Hi Alexander, Go through my research article sonkar et al 2016 Biosensors and Bioelectronics. Paper based on molecular beacon design, develop and clinical evaluation. Hope you will find your solution. If not let me know.
Best
Thanks @ Subash C Sonkar, but i did not find any information about setting PrimerBlast online tool for designing internal oligo for predesifned primer pairs.
For the same systems primer3 allow to design internal oligo, and as a rule designed probe is specific, when checking in Blast.
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