BioMasher™

Enhance Genomic DNA Extraction With The BioMasher™

The BioMasher™ Sample Preparation Device can be used on a variety of samples to create a whole cell homogenate that will serve as the starting point for a variety of DNA extraction kits and protocols. Below are examples of how to incorporate the BioMasher into several leading DNA extraction kits.

Cartagen® DNA Extraction Kit for Plant Samples
At Step 2 of the protocol:
For each sample to be processed, place ~100 to 200 mg of fresh plant tissue into the column insert, add 50 µL of Lysis Buffer (Buffer 1), homogenize the sample with the BioMasher (either by hand or attached to a lightweight handheld power drill). Remove the homogenizer and add an additional 150 µL of Lysis Buffer (Buffer 1) Replace the homogenizer and spin in a microcentrifuge for 30 seconds at 10,000 rpm. Remove and dispose of the column insert and the homogenizer and proceed with step 4 of the protocol (add 600 µL of Buffer 2 to the extract).

EPICENTRE® MasterPure™ Plant Leaf DNA Purification Kit
At Step 1 of the protocol (page 2 of the EPICENTRE MasterPure Plant Leaf DNA Purification Kit Protocol (Lit. #119):
For each sample to be processed, place ~35-100 mg of fresh weight plant leaf into the column insert, add 50 µL of Plant DNA Extraction Solution and homogenize the sample with the BioMasher (either by hand or attached to a lightweight handheld power drill). Remove the homogenizer and add an additional 150 µL of Buffer Plant DNA Extraction Solution. Replace the homogenizer and spin in a microcentrifuge for 30 seconds at 10,000 rpm. Remove and dispose of the column insert and the homogenizer. Add an additional 100 µL of Plant DNA Extraction Solution to the microcentrifuge tube containing the cell extract, and proceed with step 2 (incubate at 70ºC for 30 min.)

Invitrogen® PureLink™ Plant Total DNA Purification Kit
At Step 1 of the Invitrogen Experienced Users Procedure (page v of the PureLink Plant Total DNA Purification Kit Version A – 11 October 2004):
For each sample to be processed, place ~100 mg of fresh plant tissue into the column insert, then add 50 µL of Resuspension Buffer (R2). Homogenize the sample with the BioMasher (either by hand or attached to a lightweight handheld power drill). Remove the homogenizer and add an additional 150 µL of Resuspension Buffer (R2). Replace the homogenizer and spin in a microcentrifuge for 30 seconds at 10,000 rpm. Remove and dispose of the column insert and the homogenizer. Add 50 µL of Resuspension Buffer (R2), to the microcentrifuge tube containing the cell extract and proceed with step 3 (add 15 µL 20% SDS and 15 µL RNase A (20 mg/mL) to lysate).

Promega® Wizard® Genomic DNA Purification Kit
At Step 1 of the Promega Isolating Genomic DNA from Plant Tissue protocol (page 13 of the Wizard Genomic DNA Purification Kit Technical Manual – Revised 4/05):
For each sample to be processed, place ~100 mg of fresh plant tissue into the column insert, then add 50 µL of Nuclei Lysis Solution. Homogenize the sample with the BioMasher (either by hand or attached to a lightweight handheld power drill). Remove the homogenizer and add an additional 150 µL of Nuclei Lysis Solution. Replace the homogenizer and spin in a microcentrifuge for 30 seconds at 10,000 rpm. Remove and dispose of the column insert and the homogenizer. Add 400 µL of AP1 to the microcentrifuge tube containing the cell extract and proceed with step 2 (incubate at 65ºC for 15 minutes).

Qiagen® DNeasy® Plant Mini Kit
At Step 1 of the protocol (page 19 of the DNeasy Plant Mini and DNeasy Plant Maxi Handbook 01/2004):
For each sample to be processed, place ~100 mg of fresh plant tissue into the column insert, then add 50 µL of Buffer AP1. Homogenize the sample with the BioMasher (either by hand or attached to a lightweight handheld power drill). Remove the homogenizer and add an additonal150 µL of Buffer AP1. Replace the homogenizer and spin in a microcentrifuge for 30 seconds at 10,000RPM. Remove and dispose of the column insert and the homogenizer. Add 200 µL of AP1 to the microcentrifuge tube containing the cell lysate. Add RNase A stock solution as per the Qiagen protocol and proceed with step 2 (incubate the mixture for 10 min at 65ºC).

Other Kits and Methods
Other kits and methods can be easily adapted for use with the BioMasher. Simply pipette 50 µL of the initial lysis buffer along with the required amount of plant tissue (usually 50-200 mg) into the column insert. Homogenize the sample with the BioMasher (either by hand or attached to a lightweight handheld power drill). Remove the homogenizer and add an additonal 150 µL of the same lysis buffer. Replace the homogenizer and spin in a microcentrifuge for 30 seconds at 10,000 rpm. Remove and dispose of the column insert and the homogenizer. The total amount of lysis buffer at this point is approximately 200 µL. If the kit or method normally starts off with a larger amount, adjust the volume by adding additional lysis buffer to the cell extract so that the final volume equals the amount used in the kit or method. Then continue with the next step in the protocol.

Trademarks:

Cartagen is a trademark of Cartagen Molecular Systems Inc.

MasterPure and Epicentre are trademarks of EPICENTRE Biotechnologies.

PureLine and Invitrogen are trademarks of Invitrogen Corporation.

Wizard and Promega are trademarks of Promega Corporation.

DNeasy and Qiagen are trademarks of Qiagen GmbH.