User:Alexander S Mikheyev/Notebook/rotifer alien genes/2009/11/19

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Alien genes
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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To do

 * Oil flies
 * prepare for sequencing
 * Order:
 * GITC
 * Submit AmNat paper

PEG precipitation

 * recipe:
 * 3g PEG 8000
 * 2.19g NaCl
 * Bring up to 15ml with good water.
 * added 8ul PEG to PCR
 * spun at max speed for 15 min
 * removed PEG, added 150ul cold 80% ethanol
 * spin 7 min
 * remove ethanol, dry
 * re-suspend in 10 ul water
 * nanodrop


 * somehow I don't feel completely comfortable with potential effects of CoraLoad dye on precipitation, so I'll run a gel:

InstaGene matrix for extraction of small numbers of rotifers

 * placed 1, 2, 4 and 8 A. ricciae into 30 ul instagene buffer
 * 30 min@ 56, with one vortex, 8 min at 99C


 * 1ul and 2 ul extraction


 * 3m 94C (10s 94C; 3m 60C)x35
 * try one set with 1 ul DNA, and another set with 2 ul DNA
 * 3ul PCR product 2ul ladder
 * 1ul 4 rotifer lane -- no sample loaded into PCR