Working Sheet- qPCR, PCR, cDNA

	A	B	C	D	E	F	G	H	I	J	K	L
1	http://eagle.fish.washington.edu/trilobite/Sites_genefish_100112/Steven/Commercial%20Protocols/BioRad%20-%20Sso%20Fast%20EvaGreen%20Supermix.pdf
2		M1V1=M2V2
3		Rp_RNF66/67				Mm_flilCG1484/5
4		[Stock]	ul/rxn	Cocktail	# of NTC	ul/rxn	Cocktail	# of NTC
5	Water		7	98	2	7	98	2
6	Primer - F (uM)	10	0.5	7	# of Unknowns	0.5	7	# of Unknowns
7	Primer - R (uM)	10	0.5	7	6	0.5	7	6
8	Sso Fast EvaGreen Supermix (2X)	1X	10	140	Replicate	10	140	Replicate
9					2			2
10	cDNA		2		Total # Samples	2		Total # Samples
11	Total volume (ul)	20	20	280	14	20	280	14
12
13	Thermalcycler conditions		Cycles
14	95 C	30 sec	1				Well	Sample	Stage	Primer Set		Developmental Process
15	95 C	5 sec	30-40				A1	T1	Trocophore	Rp_RNF66_F	Rp_RNF67_R	multicellular organismal development
16	55-60 C	10 sec	30-40				A2	T2	Trocophore
17	65-95	10 sec	1				A3	V1	Veliger
18							A4	V2	Veliger
19							A5	S1	Settler
20							A6	S2	Settler
21							A7	NTC	--
22							B1	T1	Trocophore	Mm_flilCG1484_F	Mm_flilCG1485_R	nematode larval development
23							B2	T2	Trocophore
24							B3	V1	Veliger
25							B4	V2	Veliger
26							B5	S1	Settler
27							B6	S2	Settler
28							B7	NTC	--
29		PLATE SET UP
30			1	2	3	4	5	6	7
31		A	T1 Rp	T2 Rp	V1 Rp	V2 Rp	S1 Rp	S2 Rp	NTC
32		B	T1 Rp	T2 Rp	V1 Rp	V2 Rp	S1 Rp	S2 Rp	NTC
33		C	T1 Mm	T2 Mm	V1 Mm	V2 Mm	S1 Mm	S2 Mm	NTC
34		D	T1 Mm	T2 Mm	V1 Mm	V2 Mm	S1 Mm	S2 Mm	NTC
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