Materials and methods for processing blood samples
Provided herein are materials and methods relating to cell-free DNA. In particular, the technology relates to methods and materials for the preparation and handling of blood samples for future use in applications involving cell-free DNA.
1. A method for quantifying cell-free DNA, the method comprising:
(a) providing a tube comprising
an outer wall and a base defining an internal volume for containing a blood sample;
a chemical preservative disposed in the internal volume, wherein the chemical preservative comprises one or more of metaformaldehyde, paraformaldehyde, poloxamer, glycerol, propranolol, dex-propranolol, methacrylate monomer, methacrylate polymer, or bis-dienoyl phosphatidylcholine;
a physical separator disposed in the internal volume, wherein the physical separator substantially separates the blood sample into at least a plasma component and a blood cell component when the tube is centrifuged and prevents aspiration of the blood cell component during subsequent isolation of the plasma component from the tube; and
a blood sample disposed in the tube;
(b) centrifuging the tube at a first fixed speed to separate the blood sample into at least a plasma component and a blood cell component;
(c) isolating the plasma component;
(d) centrifuging the isolated plasma component at a second fixed speed to obtain a purified plasma component, wherein the first fixed speed and the second fixed speed are each independently selected from a speed ranging from 500×g to 2000×g; and
(e) quantifying cell-free DNA present in the further purified plasma component.
2. The method of claim 1 , further comprising isolating the further purified plasma component after step (d).
3. The method of claim 2 , further comprising storing the isolated further purified plasma component at a temperature of −20° C. or colder.
4. The method of claim 2 , comprising determining an amount of cf-DNA in the further purified isolated plasma component.
5. The method of claim 1 , wherein the blood cell component includes a white blood cell component and a red blood cell component.
6. The method of claim 5 , further comprising isolating the white blood cell component after step (b).
7. The method of claim 6 , wherein the method further comprises storing the isolated white blood cell component at a temperature of −20° C. or colder.
8. The method of claim 1 , wherein the first fixed speed and the second fixed speed are the same.
9. The method of claim 8 , wherein the first fixed speed and the second fixed speed are each 1100×g.
10. The method of claim 8 , wherein the first fixed speed and the second fixed speed are each 1400×g.
11. The method of claim 1 , wherein step (e) comprises quantifying cell-free DNA comprising at least one mutation.
12. The method of claim 1 , wherein step (e) comprises quantifying cell-free DNA derived from a transplant.
13. The method of claim 1 , wherein step (e) comprises quantifying cell-free DNA derived from a cancer.
14. The method of claim 1 , wherein step (e) comprises quantifying cell-free DNA derived from a fetus.