Tag Archives: 15:9-146

DNA Extraction & Quantification – Ava Withering Syndrome Transmission Study Tissues

Isolated DNA from 24 tissue samples provided by Ava. Presumably, the tissues were digestive gland and I believe they were preserved in ethanol. The list of samples can be seen in the results below.

DNA was extracted using the QIAmp Fast DNA Stool Mini Kit (Qiagen) following the manufacturer’s protocol with the following options:

  • Samples were briefly homogenized (due to their stiffness resulting from ethanol fixation) in the InhibitEX Buffer using disposable plastic pestles.
  • Homogenized tissue was incubated at 95C to maximize cell lysis
  • Followed “human DNA analysis” protocol for remainder of protocol (to maximize sample recovery)
  • Eluted DNA with 100μL Buffer ATE

After extraction, the samples were quantified using the Roberts Lab Qubit 3.0 (Life Technologies) using the Qubit ds DNA BR reagents. Used 1μL of each sample.

Samples were stored at 4C in FSH240 in the box “Ava WS Transmission DNA Extractions by Sam Box 1″. See the master spreadsheet at the bottom of this post for specific sample locations within this box.

Results:

cage_number accession_number concentration(ng/uL)
21 15:11-89 168
21 15:11-90 69.2
21 15:11-91 70.4
21 15:11-92 58.8
21 15:11-93 61.6
17 15:11-84 48
17 15:11-85 80
17 15:11-86 138
17 15:11-87 68
17 15:11-88 18.2
23 15:9-144 60
23 15:9-145 72
23 15:9-146 121
23 15:9-147 159
23 15:9-148 41.8
20 15:11-100 29