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Stir together the buttermilk and poppy seeds in a glass measuring cup or small glass bowl. Let sit for 2 hours.
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Once your poppy seeds have soaked, preheat the oven to 350 degrees. Spray 2 8-inch round cake pans with cooking spray. Line the bottoms of the pans with parchment, spray the parchment with cooking spray, and set pans aside.
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Whisk together the flour, baking powder, baking soda, and salt in a medium bowl. Set aside.
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In a large mixing bowl, beat the butter with with an electric mixer until smooth.
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Add the sugar, vanilla, lemon zest, and eggs and beat on medium until very light and fluffy, about 5 minutes. (Stop the mixer a few times to scrape the sides and bottom of the bowl.)
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With mixer on low, beat in half of the dry ingredients until just combined. Beat in half of the buttermilk mixture.
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Beat in the remaining flour, then the remaining butter milk until just incorporated. Use a rubber spatula to stir a few times to ensure everything is well mixed.
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Divide batter between the prepared cake pans and bake for 42 to 50 minutes, until a cake tester inserted into the center of the cakes comes out clean.
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Cool the cake layers in the pans for 20 minutes, then run a knife around the edge of each cake layer to loosen. Gently turn the cakes out onto a wire rack to cool completely before frosting your cake.