User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/08/04

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Inoculated Liquid culture with yeast from slants

NCYC 2827, 2878, 415, 971 and 814

1) 10mL Glucose+Yeast Media broth with sterile toothpick 2) Incubate at 30C


QIAquick PCR purification and Cycle Sequencing

Clean S. Dairenenses with QIAquick 4.9ng/ul

CS Reaction mixture conc. 1x 2
Big Dye Terminator 0.5 1
DMSO 0.1 0.2
5X Seq Buffer 5X 0.4 0.8
Primer (FWD or RVS) 5uM 0.4
Template * ~1-3ng/ul 4.6
2.0ul / RXN
*QIAquick Cleaned PCR product
Cycle Seq Program
96C 10 sec
50C 5 sec
60C 4 min
x60
MATA2 FWD and RVS
Genus species NCYC # sample
Saccharomyces dairenensis 777 4