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 | ≥10 µl |
---|---|
Equipment | Microcentrifuge, Vortex |
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, or serum from humans, mice, rats, etc. Cells from culture, buccal cells, as well as a variety of biological liquids are effectively processed using this kit. Tissue already digested with Proteinase K or mechanically homogenized. |
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 total DNA is eluted into ≥10 µl (6 µl minimum) DNA Elution Buffer or water. Human whole blood yields 1.5-3.5 µg DNA per 50 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 kit 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 |
“It was easy to work with, protocol easy to follow”
-Tinatin T.
“This kit did a good job of prepping clean genomic DNA.”
-Tara N. (United States Agricultural Research Service)
Cat # | Name | Size | Price | |
---|---|---|---|---|
D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
D3004-2-100 | g-DNA Wash Buffer | 100 ml | $30.00 | |
D3004-2-50 | g-DNA Wash Buffer | 50 ml | $18.00 | |
D3004-1-100 | Genomic Lysis Buffer | 100 ml | $60.00 | |
D3004-1-50 | Genomic Lysis Buffer | 50 ml | $34.00 | |
D3004-5-15 | DNA Pre-Wash Buffer | 15 ml | $10.00 | |
D3004-5-50 | DNA Pre-Wash Buffer | 50 ml | $26.00 | |
C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
C1004-50 | Zymo-Spin IC Columns | 50 Pack | $53.00 |
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以前先放-80再液氮,但是去-80冰箱有点远,我想能不能直接放液氮里面,我直接放过复苏成活率低了很多,有没有这种冻存盒可以直接放液氮的?
要做细胞缺氧,需要买细胞缺氧盒,但不知道哪里有卖,各位大神有没有曾经做过的,能不能告诉我在哪能购买到,哪个公司有卖。。急。。。。。。实验小白。。。。在下先在此谢过了。
心肌肌钙蛋白I,英文为CardiactroponinI,该试剂盒在POCT领域内几乎是每个厂家都必做的项目,也是多数医院科室都开展的收费项目,市场容量很大,全国一年下来保守估计有3千万人份。但有些厂家销售的特别好,有些厂家销售的很一般。这一是由于销售渠道所致、二是因为产品质量差导致。试剂盒质量差一是因为厂家自身调试工艺不太好,二是由于所选择的抗体质量不好所致。现在市面上有心肌肌钙蛋白I抗体的厂家不多,大概也就不超过10家吧,主要为进口品牌,如:HyTest、BBI、Biospacific、Medix,国产品牌为深圳菲鹏,杭州启泰。其中着重要说的是杭州启泰,这是一家成立7年的抗原、抗体研发公司,公司不大,但立足于自主研发、创新,最近新推出的cTnI抗体可以说是目前国内外最好的抗体,这也算是填补了国内该领域的空白。从事了该行业将近10年,发此贴也是真切希望国内的生物试剂类公司,都专注于研发、创新,做一些好的试剂出来。因为很多事情国内做的并不比国外差的,国内的生物同行请加油吧!
小弟现有一事儿想求助!
如下:1)凋亡的时候细胞色素c由线粒体里释放到胞浆里去的。那么如果能够测到此时在胞浆里的细胞色素c表达增多的话应该就能说明问题了
(2)可此时的问题是如果提胞浆蛋白时把线粒体也裂解了话细胞色素c也会被释放出来,还是会在胞浆里大量表达!就说不清楚是凋亡由线粒体释放的还是被人为的破坏了线粒体导致了的!!!
(3)故我的问题就是该怎么样去除这个人为的影响?怎么说明我测的胞浆里的细胞色素c就是凋亡时线粒体释放的?换句话讲就是如果能够在用细胞裂解液裂解胞浆蛋白时,将线粒体剔除掉?
是不是买个细胞色素c的测试盒就可以了啊?这个盒子有人用过吗,请指教一二!
盼复!非常感谢!
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