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- Primer designing tool - National Center for Biotechnology Information
Primer Parameters Use my own forward primer (5'->3' on plus strand) Help Clear Use my own reverse primer (5'->3' on minus strand) Help Clear PCR product size MinMax # of primers to returnPrimer melting temperatures (Tm) MinOptMaxMax Tm difference Help Primer Location Preference Prefer primers at 3' side of the templateHelp Exon intron selection A refseq mRNA sequence as PCR template input is
- Electronic-PCR (e-PCR) is retiring, use Primer-BLAST instead
Primer-BLAST automatically searches a user-selected database for your organism, or group of organisms, and it provides a nice graphical view of the primer positions
- Primer-BLAST Archives - NCBI Insights
Primer-BLAST, NCBI’s primer-designer and specificity-checker, now offers a way to help you with irrelevant off-target matches Sometimes Primer-BLAST can’t design specific primers for your target sequence because of similar non-target sequences in the database
- Primer-Blast results
Primer-BLAST was developed at NCBI to help users make primers that are specific to intended PCR target
- National Center for Biotechnology Information
The reason is that an accession or GI carries accurate information about its identity which allows primer-blast to better distinguish between intended template and off-targets Try "re-search for specific primers" feature on Primer-BLAST result page
- Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and . . .
Primer dimers can form when primers preferentially self anneal or anneal to the other primer in the reaction If this occurs, a small product of less than 100 bp will appear on the agarose gel
- Explanatory chapter: PCR primer design - PubMed
This chapter is intended as a guide on polymerase chain reaction (PCR) primer design (for information on PCR, see General PCR and Explanatory Chapter: Troubleshooting PCR) In the next section, general guidelines will be provided, followed by a discussion on primer design for specific applications …
- Primer-BLAST now designs primers for a group of related sequences
To find primers for a group of related sequences, Primer-BLAST aligns the longest sequence to the rest to find common regions It uses these to limit the locations of primers
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