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
The D3012 Zymo Quick-DNA Kits are ideal DNA isolation kits for easy, rapid isolation of total DNA (e.g., genomic, mitochondrial, viral) from a variety of biological sample sources. Whole blood (fresh or stored), buffy coat, buccal cells, cells from culture, saliva, and other biological liquid samples can be processed with these kits. Zymo-Spin Column/Plate technology enables high-quality DNA purification in minutes. PCR inhibitors are effectively removed, and the eluted DNA is ideal for PCR, nucleotide blotting, DNA sequencing, restriction endonuclease digestion, bisulfite conversion/methylation analysis, and other downstream applications.
Elution Volume
? 25 ?l
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, serum cultured 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 ?g/well total DNA is eluted into ?30 ?l DNA Elution Buffer or water. Human whole blood yields 3-7 ?g DNA per 100 ?l blood sampled. Mammalian tissues already homogenized will yield 1-3 ?g 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 | 1×106?Cells | 5-6 ?g |
| Skeletal, Heart, Lung, Brain Tissue | 1 mg | 1-3 ?g |
| Liver and Kidney Tissue | 1 mg | 3-5 ?g |
| Human Whole Blood | 100 ?l | 5-7 ?g |
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99 / 100 Bioz Stars |
The design and testing of mini-barcode markers in marine lobsters
PLoS One – Published 24 Jan 2019
‘These samples provided 67% coverage across the different families within the lobster taxonomy.. DNA from 17 species (adults and larvae combined = 19 samples) was extracted from pereiopod tissue using the Zymo Quick-DNA Universal Kit (Zymo Research), as per the manufacturer?s protocol which was modified to include an initial incubation step at 55?C overnight.. PCR reactions were 25 ?l in volume and contained 30 ng genomic DNA, 12.5 ?l OneTaq Quick-Load Master Mix (1X, BioLabs, New England), 0.50 ?l forward ‘
The TFAP2C-Regulated OCT4 Naive Enhancer Is Involved in Human Germline F…
Cell Rep – Published 22 Jan 2019
‘Distal enhancer deletion candidate lines were screened for the presence of shorter bands due to deletion.. To determine the precise mutations, genomic DNA was extracted from about 1 million cells using Quick-DNA Microprep Kit (ZYMO RESEARCH, D3021).. 1uL of the genomic DNA was used as PCR template for genotyping using Phusion High-Fidelity DNA Polymerase (NEW ENGLAND BioLabs, M0530L).’
Molecular monitoring of the diversity of human pathogenic malaria specie…
Malar J – Published 15 Jan 2019
‘HIV-1 and HIV-2 negative blood donors were analysed for the presence of possibly undetected, low-level malaria parasites using high-sensitive qPCR assays. . DNA extraction was done manually from 180 ?L whole blood using Quick-DNA Miniprep kits (Zymo Research, Irvine, USA) following manufactures? guidelines and DNA was eluted with 50 ?L Elution Buffer.. DNA samples were kept at ? 20 ?C prior to qPCR analysis.’
| Cat # | Name | Size |
|---|---|---|
| D3004-1-100 | Genomic Lysis Buffer | 100 ml |
| D3004-2-100 | g-DNA Wash Buffer | 100 ml |
| D3004-4-10 | DNA Elution Buffer | 10 ml |
| D3004-4-50 | DNA Elution Buffer | 50 ml |
| D3004-5-50 | DNA Pre-Wash Buffer | 50 ml |
| C2001 | Silicon-A Plate | 2 Plates |
| C2002 | Collection Plate | 2 Plates |
| C2003 | Elution Plate | 2 Plates |
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