Mar 09, 2019
Version 1
HPLC sample prep V.1
- 1University of Tennessee Health Science Center
Protocol Citation: Kaitly Woodard 2019. HPLC sample prep. protocols.io https://dx.doi.org/10.17504/protocols.io.yy3fxyn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 09, 2019
Last Modified: March 09, 2019
Protocol Integer ID: 21243
Abstract
Preparation of blood for hemoglobin separation (ion-exchange and reverse-phase HPLC).
Prepare 1.5mL Eppendorf tubes with 200uL of Hemolysate Reagent
Add 1uL of blood to tube, and vortex for 5 seconds.
Centrifuge at max speed for 10 minutes
Pipette out 75-100uL of supernatant into HPLC vial, careful not to disturb DNA pellet at the bottom of the tube. If pipette tip appears sticky after removal from supernatant you may have taken the DNA. Instead, disgard the sticky debris and pipette from remainder of supernatant. (The DNA and cell debris will stay together, once it is removed only desired hemolysate will remain).
Ensure vial is labeled appropriately, and no bubbles are present at the bottom of the vial. Bubbles can be removed by tapping the vial gently on the benchtop or removing by pipette.
**Reserve HPLC time on the calendar, provide sample list and samples to JJZ