The FISH-BOL collaborators' protocol

Figures & data

Figure 1.  Example of digital image associated with a DNA barcode including ruler and color bar—silver moony (Monodactylus argenteus).

Table I.  PCR and sequencing primers.

Name	Cocktail name/5′–3′ sequence	Reference
	Combinations of FishF1, F2 and FishR1, R2
FishF1	TCAACCAACCACAAAGACATTGGCAC	Ward et al. (2005)
FishF2	TCGACTAATCATAAAGATATCGGCAC	Ward et al. (2005)
FishR1	TAGACTTCTGGGTGGCCAAAGAATCA	Ward et al. (2005)
FishR2	ACTTCAGGGTGACCGAAGAATCAGAA	Ward et al. (2005)
	Fish cocktail (M13 tailed): C_FishF1t1–C_FishR1t1 (Ratio 1:1:1:1)	Ivanova et al. (2007)
VF2_t1	TGTAAAACGACGGCCAGTCAACCAACCACAAAGACATTGGCAC
FishF2_t1	TGTAAAACGACGGCCAGTCGACTAATCATAAAGATATCGGCAC
FishR2_t1	CAGGAAACAGCTATGACACTTCAGGGTGACCGAAGAATCAGAA
FR1d_t1	CAGGAAACAGCTATGACACCTCAGGGTGTCCGAARAAYCARAA
	Sequencing primers for M13-tailed PCR products
M13F	TGTAAAACGACGGCCAGT	Messing (1983)
M13R	CAGGAAACAGCTATGAC	Messing (1983)

Ward RD, Zemlak TS, Innes BH, Last PR, Hebert PDN. 2005. DNA barcoding Australia's fish species. Philos Trans R Soc B Biol Sci. 360:1847–1857.

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Ivanova NV, Zemlak TS, Hanner R, Hebert PDN. 2007. Universal primer cocktails for fish DNA barcoding. Mol Ecol Notes. 7:544–548.

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Messing J. 1983. New M13 vectors for cloning. Methods Enzymol. 101:20–78.

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