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 | ≥50 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 25 µg total DNA is eluted into ≥50 µl (30 µl minimum) 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 |
“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)
"This product was amazing! I"ve used the same type of kit (quick DNA extract) from Sigma and this was far more superior. I used the same amount of postnatal tissue as I would have for the Sigma kit, however the yield I obtained from Zymo was quite astounding considering the time of tissue digestion. Secondly, the gDNA was much "cleaner" upon measurement with our NanoDrop! "
-Stephen C. (Johns Hopkins University School of Medicine)
| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| C1011-50 | Zymo-Spin IIC Columns | 50 Pack | $55.00 | |
| C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
| D3004-5-50 | DNA Pre-Wash Buffer | 50 ml | $26.00 | |
| D3004-5-15 | DNA Pre-Wash Buffer | 15 ml | $10.00 | |
| D3004-1-50 | Genomic Lysis Buffer | 50 ml | $34.00 | |
| D3004-1-100 | Genomic Lysis Buffer | 100 ml | $60.00 | |
| D3004-2-50 | g-DNA Wash Buffer | 50 ml | $18.00 | |
| D3004-2-100 | g-DNA Wash Buffer | 100 ml | $30.00 | |
| D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 |
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生物工程的应用领域非常广泛，包括农业、工业、医学、药物学、能源、环保、冶金、化工原料等。它必将对人类社会的政治、经济、军事和生活等方面产生巨大的影响，为世界面临的资源、环境和人类健康等问题的解决提供美好的前景。展开
红细胞压积，是指红细胞占血液体积的比例，您仅仅是比参考范围略低，同时红细胞数量，和血红蛋白并没有下降，这可能是血液有所稀释所致。
白细胞计数3.9，正常的话 4-10之间，你仅仅是比参考范围略下降一丁点，本身并无意义，参考范围是大致的。而且血液还可能存在稀释可能性
中性粒细胞绝对值和百分比略低一点，同时淋巴细胞百分比略有上升，那这本身就只是略高一点点，而一般比如感冒以后，也往往可以升高，其他的红细胞分布宽度等是派生指标，没大意义的。
总之单单看这一个血常规检验，本身没有什么大的异常发现，当然也可以稍后复查，比如过2周以后复查
我是厉清，更多医疗知识，请看 厉清就是我 的新浪博客
生物工程包括五大工程，即遗传工程(基因工程)、细胞工程、微生物工程(发酵工程)、酶工程(生化工程)和生物反应器工程。在这五大领域中，前两者作用是将常规菌(或动植物细胞株)作为特定遗传物质受体，使它们获得外来基因，成为能表达超远缘性状的新物种——“工程菌”或“工程细胞株”。后三者的作用则是这一有巨大潜在价值的新物种创造良好的生长与繁殖条件，进行大规模的培养，以充分发挥其内在潜力，为人们提供巨大的经济效益 和社会效益。
生物工程的应用领域非常广泛，包括农业、工业、医学、药物学、能源、环保、冶金、化工原料等。它必将对人类社会的政治、经济、军事和生活等方面产生巨大的影响，为世界面临的资源、环境和人类健康等问题的解决提供美好的前景。展开
如题，求助。做肝癌细胞侵袭、迁移实验，用哪种细胞株比较好？
看到之前的一个朋友也发了IBC会议的PPT内容，不过好像没有这个PPT（不确定，要是重复了请见谅）。
因为会议上投影的问题，当时有些内容贾老师没介绍太多。不过内容相当不错，主要是国内比较缺乏这方面的信息和经验。可以算得上是干货吧，不过里面主要是一些规则性或方向性的内容，需要具体细节，只能花钱请她做顾问了。
AudreyJia是美籍华人，在FDA做CMCReviewer之前也一直是从事细胞开发及细胞培养的工作。她应该也是目前唯一一个生物大分子部门中，唯一从FDA离职出来从事顾问工作的。可能很多国内单抗药企大佬都认识她，可能很多园子里面的人也见过她了。
谁能赏几个积分啊？新人，没积分，一些东西想看或者下载都不行。
版主jackieustc留言:
感谢分享，鼓励新人，我给你打赏，多交流自然有收获！
Monoclonality_JIA1.pdf(952.57k)
