DNA extraction of insect tissue using the ZymoBIONICS DNA miniprep kit

Add sample to a ZR BashingBead™ Lysis Tubes (0.1 & 0.5 mm). Add 750 μl ZymoBIOMICS™ Lysis Solution to the tube and cap tightly.

Secure in a bead beater fitted with a 2 ml tube holder assembly and process using 4 repetitions of 1 minute bead beating / 3 minutes on ice.

Centrifuge the ZR BashingBead™ Lysis Tubes (0.1 & 0.5 mm) in a microcentrifuge at ≥ 10,000 x g for 1 minute.

Transfer up to 400 μl supernatant to the Zymo-Spin™ III-F Filter in a Collection Tube and centrifuge at 8,000 x g for 1 minute. Discard the Zymo-Spin™ III-F Filter.

Add 1,200 μl of ZymoBIOMICS™ DNA Binding Buffer to the filtrate in the Collection Tube from Step 4. Mix well.

Transfer 800 μl of the mixture from Step 5 to a Zymo-Spin™ IICR Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute.

Discard the flow through from the Collection Tube and repeat Step 6.

Add 400 μl ZymoBIOMICS™ DNA Wash Buffer 1 to the Zymo-Spin™ IICR Column in a new Collection Tube and centrifuge at 10,000 x g for 1 minute. Discard the flow-through.

Add 700 μl ZymoBIOMICS™ DNA Wash Buffer 2 to the Zymo-Spin™ IICR Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute. Discard the flow-through.

Add 200 μl ZymoBIOMICS™ DNA Wash Buffer 2 to the Zymo-Spin™ IICR Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute.

Transfer the Zymo-Spin™ IICR Column to a clean 1.5 ml microcentrifuge tube and add 100 μl (50 μl minimum) ZymoBIOMICS™ DNase/RNase Free Water directly to the column matrix and incubate for 1 minute. Centrifuge at 10,000 x g for 1 minute to elute the DNA.

Place a Zymo-Spin™ III-HRC Filter in a new Collection Tube and add 600 μl ZymoBIOMICS™ HRC Prep Solution. Centrifuge at 8,000 x g for 3 minutes.

Transfer the eluted DNA (Step 11) to a prepared Zymo-Spin™ III-HRC Filter in a clean 1.5 ml microcentrifuge tube and centrifuge at exactly 16,000 x g for 3 minutes.