User:Alexander S Mikheyev/Notebook/rotifer alien genes/2009/12/09

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Adineta Primers 13-18

Plate	Well	Genus	12/2/2009 Count	Sample	Extract	'	Reagents	Concentration	1X	14X
AV 10-06-09	well 6	Adineta	100+	2			H20		4.75	66.5
Boston	A1	Adineta	10+	3			10X Buffer	10X	1	14
Boston	A4	Adineta	50+	4			dye	10X	1	14
CC	A1	Adineta	subs 100+	5			dNTP	10mM	0.2	2.8
CC	A3	Adineta	subs 100+	6			primer mix	5uM	1	14
WH1	B3	Adineta	20+	7	use 12/7		Taq	5U/uM	0.05	0.7
WH1	B5	Adineta	20+	8	use 12/7		DNA	variable	2
WH1	B6	Adineta	50+	9	use 12/7		TOTAL		10	112
WH1	C4	Adineta	30+	10					Cocktail per well = 8.0ul
WH2	A6	Adineta	10+	11
WH3	A5	Adineta	20+	12					On Gels
WH3	C2	Adineta	7	13			Primers	AV10098	#1-12
								AV10092	#13-24
								AV10089	#25-36
EXTRACTION								AV10084	#37-48
InstaGene Matrix	200ul per sample							AV10082	#49-60
56C	30 min							AV10075	#61-72
99.9C	8min
							PCR	95C	3 min
								95C	10 sec
12/2/2009	5-10 individuals	12/7/2009	10-20 individuals					60C	1 min
								35 cycles
load 2.0ul 2-log ladder and 2.0ul sample 1% gel
Sample #2 Positive Control

Adineta Primers 19-24

Plate	Well	Genus	12/2/2009 Count	Sample	Extract	'	Reagents	Concentration	1X	14X
AV 10-06-09	well 6	Adineta	100+	2			H20		4.75	66.5
Boston	A1	Adineta	10+	3			10X Buffer	10X	1	14
Boston	A4	Adineta	50+	4			dye	10X	1	14
CC	A1	Adineta	subs 100+	5			dNTP	10mM	0.2	2.8
CC	A3	Adineta	subs 100+	6			primer mix	5uM	1	14
WH1	B3	Adineta	20+	7	use 12/7		Taq	5U/uM	0.05	0.7
WH1	B5	Adineta	20+	8	use 12/7		DNA	variable	2
WH1	B6	Adineta	50+	9	use 12/7		TOTAL		10	112
WH1	C4	Adineta	30+	10					Cocktail per well = 8.0ul
WH2	A6	Adineta	10+	11
WH3	A5	Adineta	20+	12					On Gels
WH3	C2	Adineta	7	13			Primers	AV10071	#1-12
								AV10045	#13-24
								AV10044	#25-36
EXTRACTION								AV10043	#37-48
InstaGene Matrix	200ul per sample							AV10042	#49-60
56C	30 min							AV10041	#61-72
99.9C	8min
							PCR	95C	3 min
								95C	10 sec
12/2/2009	5-10 individuals	12/7/2009	10-20 individuals					60C	1 min
								35 cycles
load 2.0ul 2-log ladder and 2.0ul sample 1% gel
Sample #2 Positive Control

Adineta Primers 25-30

Plate	Well	Genus	12/2/2009 Count	Sample	Extract	'	Reagents	Concentration	1X	14X
AV 10-06-09	well 6	Adineta	100+	2			H20		4.75	66.5
Boston	A1	Adineta	10+	3			10X Buffer	10X	1	14
Boston	A4	Adineta	50+	4			dye	10X	1	14
CC	A1	Adineta	subs 100+	5			dNTP	10mM	0.2	2.8
CC	A3	Adineta	subs 100+	6			primer mix	5uM	1	14
WH1	B3	Adineta	20+	7	use 12/7		Taq	5U/uM	0.05	0.7
WH1	B5	Adineta	20+	8	use 12/7		DNA	variable	2
WH1	B6	Adineta	50+	9	use 12/7		TOTAL		10	112
WH1	C4	Adineta	30+	10					Cocktail per well = 8.0ul
WH2	A6	Adineta	10+	11
WH3	A5	Adineta	20+	12					On Gels
WH3	C2	Adineta	7	13			Primers	AV10034	#1-12
								AV10028	#13-24
								AV10027	#25-36
EXTRACTION								AV10025	#37-48
InstaGene Matrix	200ul per sample							AV10016	#49-60
56C	30 min							AV10011	#61-72
99.9C	8min
							PCR	95C	3 min
								95C	10 sec
12/2/2009	5-10 individuals	12/7/2009	10-20 individuals					60C	2 min
								35 cycles
load 2.0ul 2-log ladder and 2.0ul sample 1% gel
Sample #2 Positive Control

Reloaded same PCR products on cycler - lots of primer dimers?

RERUN PCR on 14Dec09