Hair, Feather, and Nail Samples:
1. To ≤ 25 mg sample, add 90 µl Water, 90 µl Solid Tissue Buffer (Blue)
(Cat. No. D4068-2-6), 10 µl 1M DTT, and 10 µl Proteinase K (Cat.
No. D3001-2-5) in a microcentrifuge tube.
2. Mix thoroughly or vortex 10-15 seconds and then incubate the tube at
55
º
C overnight.
3. Transfer lysate to a ZR BashingBead
™
Lysis Tube (0.1 & 0.5 mm)
and then add 750 µl ZymoBIOMICS™ Lysis Solution.
4. Continue to Step 2 (page 6) and proceed with the remaining protocol
as written.
Tissue and Insect Samples
Tissue and Insect samples can be processed three different ways,
depending on the sample type and the equipment available. The
recommendations are listed next to the options below:
(A) Proteinase K - Tissue
1. Add up to 15 mg of tissue to a 1.5 ml microcentrifuge tube, then add a
solution of 95 µl water, 95 µl Solid Tissue Buffer (Blue) (Cat. No. D4068-2-
6) and 10 µl Proteinase K (Cat. No. D3001-2-5). Incubate for at least 1 hour
at 55° C or until tissue clarifies (samples can be incubated overnight without
affecting DNA quality).
2. Transfer digestion to a ZR BashingBead™ Lysis Tube (0.1 & 0.5 mm) and
add 750 µl of ZymoBIOMICS™ Lysis Solution.
3. Proceed to Step 2 (page 6) and continue with the protocol as written.
(B) Bead beating - Tissue and Insect
1. Add up to 15 mg of tissue/insect sample in a ZR BashingBead
™
Lysis Tube
(2.0 mm) (Cat. No. S6003-50) with 750 µl of ZymoBIOMICS
™
Lysis
Solution.
2. Secure in a bead beater fitted with a 2 ml tube holder assembly and process
at maximum speed for ≥ 5 minutes.
Note: Processing time will vary based on sample input and bead beater. Times
may be as little as 5 minutes when using high-speed cell disrupters (FastPrep
®
-
24) or as long as 20 minutes when using lower speeds (e.g., Vortex Genie
®
).
3. Transfer the entire lysate to the ZR BashingBead™ Lysis Tube (0.1 & 0.5
mm), proceed to Step 2 (page 6), and continue with protocol as written.
(C) Mortar & Pestle - Tissue and Insect
1. Homogenize up to 15 mg tissue/insect sample with a mortar and pestle while
submersed in liquid nitrogen.
2. Transfer the entire sample into the ZR BashingBead™ Lysis Tube (0.1 &
0.5 mm) and add 750 µl of ZymoBIOMICS™ Lysis Solution.
3. Proceed to Step 2 (page 6) and continue with the protocol as written.