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Standard Poly(A)+ Buffer Recipe
This formulation creates a buffer containing 100 ng/μL Poly(A) and 0.01% Pluronic F-68 in DNA Suspension Buffer (e.g., 10 mM Tris-HCl, pH 8.0).
Reagents Needed:
- DNA Suspension Buffer (10 mM Tris-HCl, pH 8.0)
- Poly(A) salt (e.g., potassium or sodium salt)
- Pluronic F-68 (10% solution)
Preparation Steps:
- Make a Poly(A) Stock Solution: Resuspend 100 mg of Poly(A) in 2 mL of DNA Suspension Buffer to achieve a 50 mg/mL concentration. Vortex thoroughly and store at 4°C.
- Make the Final Buffer: To 250 mL of DNA Suspension Buffer, add 0.5 mL of the 50 mg/mL Poly(A) stock solution and 0.25 mL of 10% Pluronic F-68.
- Mix & Store: Mix well. This final Poly(A)+ buffer solution can be stored at 4°C.
Why is this buffer used?
AAV vectors have a tendency to stick to standard plastic and glass surfaces, especially at very low concentrations. The Poly(A) acts as a carrier nucleic acid to prevent sample loss and non-specific adsorption, while the surfactant (Pluronic F-68) reduces surface tension, further minimizing adherence and improving droplet consistency in your ddPCR assays.
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