D6016 Zymo Quick-DNA Tissue/Insect Miniprep Kit in USA and Canada
The D6016 Zymo Quick-DNA Tissue/Insect Miniprep Kit is an insect DNA extraction kit designed for the simple and rapid isolation of DNA (e.g., genomic, viral, mitochondrial) from fresh, frozen, or stored insect specimens including mosquitoes, bees, lice, ticks, and D. melanogaster . The procedure is easy and can be completed in minutes: samples are rapidly and efficiently lysed by bead beating with our state of the art, ultra-high density BashingBeads. The DNA is then isolated and purified using our Zymo-Spin column technology and is ideal for downstream molecular-based applications including PCR, array, genotyping, etc. The procedure is compatible with mammalian tissues, whole blood, and cultured cells.
|Applicable For||All sensitive downstream applications such as qPCR and Next-Generation Sequencing.|
|Elution Volume||≥ 35 µl|
|Equipment||Microcentrifuge, vortex, cell disruptor/pulverizer|
|Purity||A260/A280 nm ≥1.8.|
|Sample Source||Small amounts (n ≥ 1 and ≤ 50 mg) of fresh, frozen, or stored insects. Also, compatible with fresh or frozen mammalian tissues, as well as cultured cells and whole blood.|
|Sample Storage||DNA stored at ≤ -20°C.|
|Size Range||Capable of recovering genomic DNA up to and above 40 kb. Typical fragment sizes range from 25 kb – 35 kb. If present, parasitic and viral DNA will also be recovered.|
|Yield||≤ 25 µg total DNA|
Q1: My lysate seems viscous. What is causing this to happen? How can I fix this?
A viscous sample can indicate incomplete sample lysis. Try using less of your sample and optimize bead beating conditions (duration, speed, time) to ensure samples are thoroughly lysed. After bead beating, pellet the cell debris before moving on. Adding more Genomic Lysis buffer to the lysate can help dilute and deproteinate the sample, making the sample less viscous and more suitable for DNA recovery.
Q2: Are there any tips in optimzing bead beating conditions?
We have validated our kits with both high-speed homgenizers and low-speed disruptors. We highly recommend users to optimize their bead beating conditions for their own instruments. We recommend using a 2 ml-tube adapter to ensure that the bead beating is efficent (do not use adapters made of foam). For high-speed homogenizers, we recommend a total of 5 mins bead beating (1 min interval at 6.5 m/s with 5 mins rest, repeat 5 times). For low-speed cell disruptors, we recommend 30 mins at max speed.
Q3: Is it necessary to add beta-mercaptoethanol? Can this step be substituted or omitted?
Addition of beta-mercaptoethanol is recommended to enhance sample lysis, but can be substituted with dithiothreitol (DTT, final concentration of 10 mM). However, if bead beating is optimized and lysis is efficient, the addition of BME is not necessary and can be omitted.
Q4: When can an RNase A treatment be implemented in the protocol?
No additional RNase A treatment is required when processing samples within kit capacity. The selective chemistry allows for binding of double stranded DNA to the column and for RNA to flow through.