Finished re-extraction of the 5 1800s specimens using the Qiagen DNEasy Kit. Digestions began 09/01/25 7pm, pulled digestions 09/04/25 ~7am. See digestion details here.
Followed Qiagen DNEasy extraction protocol found here.
Samples: 50368, 50603, 19054, 14399, 42137.
Notes:
Eluted in 100uL Qiagen AE
50368 has lots of big sclerites.
After Qiagen extraction, immediately cleaned extractions using the Zymo Genomic DNA Clean and Concentrate Kit. Followed protocol included here.
Eluted in 50uL of 60C Zymo EB/
Notes:
- 42137 had an almost black Qiagen extraction, but almost clear after Zymo!
Qubit of Zymo-cleaned extractions:
Used dsDNA 1X HS kit, followed protocol found here, using 1uL of sample input.
Standard 1: 35.68
Standard 2: 16857.03
50368: 84.0 ng/uL
50603: 9.72 ng/uL
19054: 1.58 ng/uL
14399: 30.8 ng/uL
42137: 54.0 ng/uL
Next step would normally be a gel, but I’m short on time and want to conserve DNA. Since I already know the KAPA bead-to-sample ratio I want to use, I’ll go straight to bead clean/size selection.