Primer Evaluation
Primer Sequences
>MiFsh-U_F-a
GCCGGTAAAACTCGTGCCAGC
>MiFish-U_R
CATAGTGGGGTATCTAATCCCAGTTTG
Expected Amplicon Size
MIDORI
MIDORI is a reference database of DNA sequences, which can be used for taxonomic assignments of all eukaryota mitochondrial DNA sequences.
In silico PCR Hits:
N(total) = 132,578
N(PCR-e0) = 4,649
N(PCR-e1) = 6,222
N(PCR-e2) = 9,647
N(PCR-e3) = 15,387
☛ e: maximum number of mis-matches per primer
Taxa Composition:
Fish
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N(Actinopteri) = 7,355
N(Cladistia) = 44
N(Chondrichthyes) = 191
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N(Fish) = 7,590
🐟 Fish List Actinopteri 🐟 Fish List Cladistia 🐟 Fish List Chondrichthyes
Not-Fish
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N(Amphibia) = 1,981
N(Aves) = 690
N(Dipnoi) = 3
N(Hyperoartia) = 14
N(Insecta) = 3
N(Lepidosauria) = 2,911
N(Mammalia) = 2,112
N(UnknownClass) = 83
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N(Not-Fish) = 7,797
Not-🐟 Fish List
⇒ Expected 12S fragment size: Amplicon (219nt) - Primer-F (21nt) - Primer-R (27nt) = 171nt