User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/07/29

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Chelex Yeast Extraction

1) Pellet 25.0ul of growing culture (1min at 15,000rpm) 2) Remove supernatent 3) Add 100.0ul of Chelex 4) Vortex and spin down 5) Boil for 10min 6) 1 min at 27C (to prevent lids from popping when opening) 7) Vortex and spin down at 15,000rpm for 1 min before use

Genus species NCYC # Yeast Culture Number
Saccharomyces bayanus 415 13
Saccharomyces exiguus 814 14
Saccharomyces unisporus 971 15
Saccharomyces rosinii 2827 16
Saccharomyces barnettii 2878 17

MATA2 Yeast Amplification

EX Takara RXN Mixture conc 1X 8
Milli Q water 6.15 49.2
Takara EX Buffer 10X 1 8
dNTP 2.5uM 0.8 6.4
Primer Mix 5uM 1 8
EX Taq 5U 0.05 0.4
Template Chelex Extraction 1
72
9.0ul / RXN and 1.0ul Chelex Extraction
Genus species NCYC # sample
Saccharomyces bayanus 415 13
Saccharomyces exiguus 814 14
Saccharomyces unisporus 971 15
Saccharomyces rosinii 2827 16
Saccharomyces barnettii 2878 17
Saccharomyces cerevisiae 1001 (+)
Negative Control (-)
PCR cycle
98C 10 sec
60C 1 min
x15

Load 2.0ul sample

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