La Biblioteca Nacional cuenta con varias Salas de Lectura:
Colección Cubana "Antonio Bachiller y Morales"
- Colección Cubana
- Colección de Grabados Cubanos y Extranjeros
- Fondos Raros y Valiosos
- Fototeca
- Manuscritos
- Mapoteca
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Sala de arte "Wilfredo Lam"
- Arte
- Carteles
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Sala de Mùsica "Leòn-Murguecia"
- Música
- Programas de mano
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Mediateca
- Materiales audiovisuales y en formato digital
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Sala de Etnología y Folclore "Fernando Ortiz"
- Libros y Publicaciones relacionados con la Etnología y Folklore
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Sala Rusa "Alexander Pushkin"
- Documentos en idioma ruso
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Sala para la atención a personas con necesidades especiales "Frank Emilio"
- Colección de documentos en Braille y libros hablados
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Sala de Referencia "Leonor Pérez"
- Referencias (presenciales, por teléfono e email y virtuales a través del Portal institucional)
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Sala de Servicios Generales "Domingo Figarola Caneda"
- Archivo de la Biblioteca Nacional
- Colección de documentos Siglos XX y XXI
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Sala Circulante María Teresa Freyre de Andrade"
- Servicios de préstamos internos y externos de las colecciones para adultos.
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