Bullet_Blender_Homogenizer_Protocol_Homogenization_Blueberry_Vaccinium
0 likes137 views
This document provides a protocol for homogenizing blueberries using the Bullet Blender. The protocol calls for sectioning blueberries into quarters, adding stainless steel beads at 1.5 times the mass of fruit, and buffer at 2 volumes per mass of sample. Samples are blended at speed 8 for 3 minutes, inspected, and blended again if needed before proceeding with downstream applications such as nucleic acid isolation or protein extraction. Safety precautions are noted for balancing tubes during centrifugation.
Bullet_Blender_Homogenizer_Protocol_Homogenization_Blueberry_Vaccinium
- 1. Protocol for Blueberry Homogenization in the Bullet Blender™ The protocol described in this document is for the use of the Bullet Blender™ for the homogenization of blueberry (flesh, seeds and skin from the genus Vaccinium L.). This protocol does not specify a particular buffer - you may choose which is most appropriate for your downstream application (nucleic acid isolation, protein extraction, etc.). Materials Required: blueberry, saline, aspirator, Bullet Blender™, homogenization buffer, pipettor, microcentrifuge tubes, 0.9-2.0mm stainless steel bead blend (part number SSB14B) Instructions 1. OPTIONAL: Wash blueberry 3x with ~1mL PBS. Aspirate. NOTE: This step removes some external contaminants and debris. 2. Section blueberry into quarters. Place quarter (100-200mg) into a microcentrifuge tube. Size may vary depending on species. 3. Add a mass of the stainless steel bead blend equal to 1.5X the mass of fruit. One scoop of beads ≈ 200mg. 4. Add 0.2ml to 0.6ml buffer, i.e. 2 volumes of buffer to the tube for every mass of sample. 5. Close the microcentrifuge tubes. 6. Place tubes into the Bullet Blender™. 7. Set controls for SPEED 8 and TIME 3 minutes. Press Start. 8. After the run, remove tubes from the instrument. 9. Visually inspect samples. If homogenization is unsatisfactory, run for another three minutes at the SPEED 10. 10. Remove sample tubes from the Bullet Blender™ and proceed with your downstream application. SAFETY NOTE!!! When using a centrifuge to separate your homogenate from the debris and beads, make sure your tubes are balanced. before after Date 08/06/2009 Next Advance, Inc. 24 Prospect Avenue, Averill Park, NY 12018 USA Phone (518) 674-3510 Fax (518) 674-0189 www.nextadvance.com