Horseshoe Crab DNA Quantification - Qubit Assay Broad Range dsDNA

Date Performed: September 2nd, 2020
Tissue samples quantified: 425, 444, 445, 491, 818, 824, 887

Quantification Set-up for Quibit

1. Retrieved samples for quantification from -20 freezer and placed in ice bucket
2. Retrieved standards for quantification from 4 degree refrigerator to come to room temp
3. Retrieved Quant-IT Reagent and Quibit Buffer from Puritz bench
4. Calculated master-mix formula
   • n = number of samples + number of standards(2) + 1 (for error)
   • master mix = n ul of reagent + 199*n ul of buffer
5. Mixed master mix components in 5mL tube
6. Vortexed tube then spun in large centrifuge
7. Set up thin walled Axygen tubes for each sample and standards
8. Added 190ul of master mix to tubes for standards
9. Added 199ul of master mix to tubes for Samples
10. Vortexed standards and added 10ul of each to corresponding Axygen tube
11. Vortexed and spun down samples
12. Carefully added 1ul of each sample to corresponding Axygen tube. Made sure to watch 1ul come out of pipette to ensure it did not stick to the side/not come out
13. Vortexed and spun all Axygen tubes at Qubit station before reading

Reading Samples

1. Woke up Quibit and selected Broad Range DNA assay
2. Read standards to ensure 10 fold difference between first and second standard
3. Read each sample twice