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
09dec09 primer13-18 1-38 text.jpg
09dec09 primer13-18 39-60 text.jpg
09dec09 primer13-18 61-72 text.jpg

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
09dec09 primer19-24 1-38 text.jpg
09dec09 primer19-24 39-60 text.jpg
09dec09 primer19-24 61-72 text.jpg

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