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 |
ebiomall.com
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希望有大神赐教!谢谢！
G418浓度之前师兄已经摸索出来，这次筛选直接用师兄浓度。
1、6孔板转染后多长时间开始进行G418筛选？
2、G418筛选是在6孔板中继续筛选还是转到24/96孔？大皿？
3、G418浓度什么时候减量
4、？筛选过程中荧光观察一小部分克隆不错，如何挑选出来？
5、整个筛选过程大约需要多长时间？
lipo3000筛选
谢谢大家！
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
你的第一部分，白细胞和中性粒细胞偏低，说明免疫力低，最近要多保养，不受凉，近期容易生病。不过数据也不是太低的，也不用过于担心，多吃多睡，过一段时间复查，可能会好转为正常。另外消炎药，止痛，退热药尽量不吃，这些药会加重白细胞下降的。
第二部分，最主要的红细胞和血红蛋白是正常的，总体看还是正常的。但是下面数据有点低，而且只低一点，问题不大，多吃点米饭，红枣，肉等，会好转的。
第三部分，血小板部分是正常的。
你这种检查单，最主要的是免疫力低下，注意多吃多睡，别受凉，少烦神少劳累，引起白细胞低的药物尽量不吃。
