Tag Archives: gray whale

gDNA Isolation

Isolated gDNA from gray whale skin, human cheek cells (my own!) and two different species of algae (species 1, species 2) using Qiagen’s DNEasy Blood & Tissue Kit according to protocol. Incubated all samples at 55C for 1hr. Eluted DNA with 50uL of Buffer AE. Spec’d samples on NanoDrop 1000.

Cheek cells were scraped from the inside of my cheek with a sterile toothpick. The toothpick was transferred to a 1.5mL snap cap tube containing the appropriate buffer. The tube was vortexed to help dislodge cells from the toothpick. The toothpick was then removed and the sample treated according to protocol.

1mL of algae cells were collected from each liquid culture, cells were pelleted by spinning 16,000g for 1min @ RT, supe removed, 180uL of Buffer ATL added and then vortexed to dislodge/break up pellet. Sample was treated according to protocol.

Results:

Well, no detectable quantities of DNA in 3 of the 4 samples. There appears to be something in the gray whale skin gDNA extraction, however the OD260/280 ratio is just crazy, leading me to believe that there’s not really any usable DNA present. Will give gray whale gDNA sample to Caroline for class and will talk with Steven and Caroline concerning their interest in performing another quick extraction on more algae to use for class this afternoon.

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EtOH Precipitation – Whale gDNA from 20101022

Precipitated whale gDNA in hopes of producing a sample with a higher concentration. Added 0.1 vols of 3M NaOAc (10uL), 2.5 vols of 100% EtOH (275uL), mixed thoroughly and incubated @ -20C for 1hr. DNA was pelleted by spinning sample @ 16,000g, 30mins, 4C. No visible pellet. Supe was removed, sample was washed with 1mL 75% EtOH, and pelleted by spinning @ 16,000g, 15mins, 4C. Supe was removed, sample resuspended in 10uL nuclease-free H2O and spec’d.

Results:

No DNA to speak of (spec data not shown).

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gDNA Isolation

Isolated gDNA from crab (unknown species), starfish exposed to RoundUp (unknown species) and gray whale blubber using Qiagen’s DNEasy Kit, according to manufacturer’s protocol. Tissue was incubated at 55C with Proteinase K for 1hr. gDNA was eluted with 100uL of Buffer AE and spec’d.

Results:

Gray whale sample has virtually no DNA. Will try to precipitate the whale gDNA in order to obtain a more concentrated sample. The other two samples look good, with good yields and good OD260/280 ratios.

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