Table 2. Results of experiment 1a examining the effect of incubation time in the T1 buffer with proteinase K (from the Nucleospin DNA extraction kit) on non-destructive total genomic extractions of species from nine invertebrate groups.

The success of non-destructive (ND) extraction (Extraction 1) for standard DNA barcoding PCR is indicated by the ‘ +’ symbol (‘−’ for no PCR product), while the success of the second Chelex extraction (Extraction 2) on the same material is indicated the adjacent column. GenBank accession numbers for sequenced specimens are given in parenthesis.

Order/ subclass	Species (family)	Sclerotization	Life stage	Incubation time for Extraction 1
				30 min		1 hr		3 hr
				Extraction 1 (1st ND)	Extraction 2 (2nd Chelex)	Extraction 1 (1st ND)	Extraction 2 (2nd Chelex)	Extraction 1 (1st ND)	Extraction 2 (2nd Chelex)
Oligochaeta	Lumbriculus variegatus (Lumbriculidae)	Soft bodied	Adult	+ (MG976202)	–	+	–
Gastropoda	Physa acuta (Physidae)	Soft bodied/shell	Adult	+ (MG976201)	+ (MG976114)	+	+	+	–
Amphipoda	Austrochiltonia subtenuis (Chiltoniidae)	Light sclerotized	Adult	+ (MG976104)	+ (MG976105)	+	+
Diptera	Procladius villosimanus (Chironomidae)	Sclerotized head	Larvae			+ (MG976143)	+ (MG976144)	+	+
Trichoptera	Hellyethira simplex (Hydroptilidae)	Sclerotized head and thorax	Larvae	+ (MG976170)	+ (MG976171)	+ (+)
Ephemeroptera	Offadens sp. (Baetidae)	Sclerotized	Nymph	+ (MG976108)	+ (MG976109)	+ (+)
Plecoptera	Dinotoperla thwaites (Gripopterygidae)	Sclerotized	Nymph	+ (MG976167)	+ (MG976167)	+ (+)
Hemiptera	Micronecta sp. (Corixidae)	Sclerotized	Adult	+ (MG976178)	+ (MG976178)	+ (+)
Coleoptera	Necterosoma sp. (Dytiscidae)	Heavily sclerotized	Adult	–	+	+ (MH000193)	+a (MH000194)	+	+a

Notes.

^aExtraction 2 using crushed legs failed, but 2 hr incubation of whole animal in Chelex (with proteinase K) was successful.