Highlights
- Easy purification of high-quality DNA from whole blood, buffy coat, swabs, or cultured cells.
- Protocol excludes the use of Proteinase K and organic denaturants for biofluid and cell samples.
- Eluted, inhibitor-free DNA is ideal for PCR, endonuclease digestion, bisulfite conversion/methylation detection, sequencing, genotyping, etc.
Description
| Elution Volume | ≥ 25 µl |
|---|---|
| Equipment | Microcentrifuge, vortex, centrifuge w/ microplate carriers |
| Purity | High-quality DNA is eluted with DNA Elution Buffer or water. DNA is especially well suited for PCR and other downstream applications. A260/A280>1.8 |
| Sample Source | Whole blood, plasma, serumcultured cells, buccal cells, tissue already digested with Proteinase K or mechanically homogenized from human, mouse, rat, etc. are effectively processed using this kit. |
| Size Range | Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered |
| Workflow | Unique lysis buffer system omits the need for Proteinase K digestion for biological fluids and cell culture samples. |
| Yield | Up to 5 µg/well total DNA is eluted into ≥30 µl DNA Elution Buffer or water. Human whole blood yields 3-7 µg DNA per 100 µl blood sampled. Mammalian tissues already homogenized will yield 1-3 µg DNA per mg. |
Q1: What is the difference between Quick-DNA and Quick-DNA Plus kits?
The Quick-DNA is optimized for cells, soft tissues, and homogenized/digested samples using a single lysis/binding buffer. The Quick-DNA Plus kits contain an optimized Proteinase K for processing a wider variety of sample inputs, such as cells, blood, tissues, etc. The upgraded Quick-DNA Plus typically recovers more DNA.
Q2: I’m seeing some yield inconsistencies with my blood samples, what’s happening?
White blood cells, which are the major source of genomic DNA in blood, easily and quickly settles. Mix the blood sample well prior to taking an aliquot for purification.
Q3: Can the Quick-DNA kit be used with bacterial samples?
E. coli cells are easy-to-lyse and can be processed directly. For other microbes, additional pretreatment (e.g. enzymatic digestion or mechanical lysis) can be implemented and then processed with the Quick-DNA Kit. Alternatively, for an all-inclusive kit to process all microbes, use any of Zymo Research’s Environmental Kits (e.g. Quick-DNA Fungal/Bacterial, Quick-DNA Fecal/Soil, ZymoBIOMICS DNA, etc.) for DNA isolation.
Q4: Can I use the Quick-DNA kit to clean-up previously isolated DNA?
No, the kit is designed for direct use with biological samples. For clean-up of isolated DNA, please use the Genomic DNA Clean & Concentrator or the DNA Clean & Concentrator kits.
Q5: Can Quick-DNA process crude lysates?
Yes, add 4 volumes of Genomic Lysis Buffer to 1 volume of crude lysate, homogenized, or digested sample (see Cell Suspensions and Proteinase K Digested Samples) and proceed with the remainder of the protocol.
Q6: What is the purpose of adding beta-mercaptoethanol? Can this step be substituted or omitted?
Beta-mercaptoethanol is a reducing agent that helps break down proteins and improves DNA recovery and purity. Addition of beta-mercaptoethanol is recommended to enhance sample lysis, but can be substituted with dithiothreitol (DTT, final concentration of 10 mM) or omitted.
Q7: Is it possible to add an RNase A treatment to the protocol?
The Quick-DNA kits recover RNA-free genomic DNA. The selective chemistry allows for binding of double stranded DNA to the column and for RNA to flow through. No RNase A treatment is required when processing samples within kit specifications.
Q8: What are the expected yields for each sample type?
Keep in mind that there is sample to sample variability.
| Sample Type | Input Amount | Expected Yield |
|---|---|---|
| Eukaryotic Cells | 1x106 Cells | 5-6 µg |
| Skeletal, Heart, Lung, Brain Tissue | 1 mg | 1-3 µg |
| Liver and Kidney Tissue | 1 mg | 3-5 µg |
| Human Whole Blood | 100 µl | 5-7 µg |
| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| D3004-4-50 | DNA Elution Buffer | 50 ml | $32.00 | |
| D3004-5-50 | DNA Pre-Wash Buffer | 50 ml | $26.00 | |
| D3004-1-100 | Genomic Lysis Buffer | 100 ml | $60.00 | |
| D3004-2-100 | g-DNA Wash Buffer | 100 ml | $30.00 | |
| D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
| C2003 | Elution Plate | 2 Plates | $19.00 | |
| C2002 | Collection Plate | 2 Plates | $22.00 | |
| C2001 | Silicon-A Plate | 2 Plates | $129.00 |
