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

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Contents

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
Saccharomycesbayanus41513
Saccharomycesexiguus81414
Saccharomycesunisporus97115
Saccharomycesrosinii282716
Saccharomycesbarnettii287817

MATA2 Yeast Amplification

EX Takara RXN Mixture conc 1X 8
Milli Q water6.1549.2
Takara EX Buffer10X18
dNTP2.5uM0.86.4
Primer Mix5uM18
EX Taq5U0.050.4
TemplateChelex Extraction1
72
9.0ul / RXN and 1.0ul Chelex Extraction
GenusspeciesNCYC #sample
Saccharomycesbayanus41513
Saccharomycesexiguus81414
Saccharomycesunisporus97115
Saccharomycesrosinii282716
Saccharomycesbarnettii287817
Saccharomycescerevisiae1001(+)
Negative Control(-)
PCR cycle
98C10 sec
60C1 min
x15

Load 2.0ul sample

Image:29July2010.jpg

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