Highlights
- Rapid method for the isolation of inhibitor-free, PCR-quality DNA from fecal samples in minutes, including those from humans, birds, rats, mice, cattle, etc.
- Ultra-high density BashingBeads are fracture resistant and chemically inert.
- Zymo-Spin column and unique filtration technologies effectively removes PCR inhibitors from the DNA product.
Description
Applicable For | All sensitive downstream applications such as qPCR and Next-Generation Sequencing. |
---|---|
Elution Volume | ≥ 20 µl |
Equipment | Microcentrifuge, vortex, cell disruptor/pulverizer |
Processing Volume | ≤50 mg feces, ≤ 250 mg soil, ≤20 mg fungal/bacterial cells (wet weight) |
Purity | A260/A280 nm ≥1.8. |
Sample Source | Feces or soil |
Sample Storage | DNA stored at ≤ -20°C. |
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. |
Type | Total DNA |
Yield | ≤ 5 µg total DNA |
Q1: My lysate seems viscous. What is causing this to happen? How can I fix this?
A viscous sample can indicate incomplete sample lysis. Try using less of your sample and optimize bead beating conditions (duration, speed, time) to ensure samples are thoroughly lysed. After bead beating, pellet the cell debris before moving on. Adding more Genomic Lysis buffer to the lysate can help dilute and deproteinate the sample, making the sample less viscous and more suitable for DNA recovery.
Q2: Are there any tips in optimzing bead beating conditions?
We have validated our kits with both high-speed homgenizers and low-speed disruptors. We highly recommend users to optimize their bead beating conditions for their own instruments. We recommend using a 2 ml-tube adapter to ensure that the bead beating is efficent (do not use adapters made of foam). For high-speed homogenizers, we recommend a total of 5 mins bead beating (1 min interval at 6.5 m/s with 5 mins rest, repeat 5 times). For low-speed cell disruptors, we recommend 30 mins at max speed.
Q3: What is the purpose of Zymo-Spin II-µHRC step?
Environmental samples often contain inhibitors such as polyphenolics, humic/fulvic acids, tannins, melanins, etc. that affect downstream applications such as PCR. Once the DNA is eluted off the binding column, the DNA is then passed through the Zymo-Spin II-µHRC to remove the PCR inhibitors, and the DNA is then ready for downstream applications. The Zymo-Spin II-µHRC does not bind DNA, it simply removes the PCR inhibitors.
Q4: Is it necessary to add beta-mercaptoethanol? Can this step be substituted or omitted?
Addition of beta-mercaptoethanol is recommended to enhance sample lysis, but can be substituted with dithiothreitol (DTT, final concentration of 10 mM). However, if bead beating is optimized and lysis is efficient, the addition of BME is not necessary and can be omitted.
Q5: When can an RNase A treatment be implemented in the protocol?
No additional RNase A treatment is required when processing samples within kit capacity. The selective chemistry allows for binding of double stranded DNA to the column and for RNA to flow through.
Cat # | Name | Size | Price | |
---|---|---|---|---|
D3004-1-100 | Genomic Lysis Buffer | 100 ml | $60.00 | |
D3004-2-50 | g-DNA Wash Buffer | 50 ml | $18.00 | |
D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
D3004-5-15 | DNA Pre-Wash Buffer | 15 ml | $10.00 | |
D6001-3-40 | BashingBead Buffer | 40 ml | $29.00 | |
D6035-1-30 | Prep Solution | 30 ml | $18.00 | |
C1057-50 | Zymo-Spin III-F Filters | 50 Pack | $59.00 | |
C1004-50 | Zymo-Spin IC Columns | 50 Pack | $53.00 | |
C1059-50 | Zymo-Spin II-µHRC Filters | 50 Pack | $108.00 | |
S6012-50 | ZR BashingBead Lysis Tubes (0.1 & 0.5 mm) | 50 Tubes | $101.00 |
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请各位大侠给予帮助!!
谢谢!!
取片剂一片,照溶出度测定法(中国药典2000年版二部附录ⅩC第三法),以水250ml为溶剂,转速为每分钟50转,依法操作,分别经15、30、45、60、75、210分钟取溶液滤过,精密量取续滤液5ml于分液漏斗中,加入pH7.4磷酸盐缓冲液5ml,5ml0.3%溴麝香草酚蓝溶液,用15ml氯仿分两次萃取,合并萃取液,加入0.4g无水硫酸钠,照分光光度法(中国药典2000年版二部附录ⅥA),在410nm波长处测得溶出A值。现15、30、45、60、75、210分钟溶出A值分别为0.0413、0.0544、0.0437、0.0479、0.0394、0.0302。(测定吸收度偏小是否不准,有影响。)
请各位站友指教。
健那绿染液是一种活体染液,实验对象必须是活细胞,健那绿可以使活细胞中的线粒体呈现蓝绿色,而细胞质接近无色。
而且样品中的无水硫酸钠未变色,而做标准曲线的五个和空白对照的变为蓝色了,请高手指教,多谢!
还有,是否变蓝对测定结果有影响吗?
谢了哈
欢迎你!请下次规范发贴:)
产品主要应用:点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
氨基类染料是包含自由氨基的活性染料,染料可与活化羧酸衍生物和其他亲电子的试剂结合。比如:氨基与EDC-活化的羧基结合。
相关产品如下:
中文名英文名产品编号分子结构Cy7.5胺Cy7.5amineAGF1350A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110804_5250.jpg[/img]Cy5胺Cy5amineAGF1332A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110715_7750.jpg[/img]相似系列产品:
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产品主要应用:点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
菁染料是性能优良的荧光标记染料,摩尔吸光系数在荧光染料中是最高的。N-羟基琥珀酰亚胺酯是最常用的脂肪氨基标记试剂,广泛用于蛋白质、氨基酸多肽、抗体、核酸及其他生物分子的标记和检测。通过改变次甲基链的长度,可改变其荧光发射波长,每增加一个双键,按照Huoffman规则正好红移约100nm。
菁染料Cy3和Cy5已成为基因芯片的首选荧光标记物;另外,Cy5,Cy5.5和Cy7,Cy7.5的吸收在近红外区背景非常低,是荧光强度最高、最稳定的长波长染料,特别适合于活体小动物体内成像。但由于菁染料,尤其是不对称菁染料的合成副反应多,副产物极性相近,产物的分离提纯相当困难。菁染料特别是水溶性菁染料分子极性大,分离提纯越加困难。Lumiprobe供应脂溶性和水溶性菁染料。
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Sulfo-Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704104047_6968.jpg[/img]相似系列产品:
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甲基绿派洛宁
还有一种忘了。
假期没法联系老师呵呵。
这三种的作用都是什么。
健那绿——高中唯一一个活体染色剂。染线粒体的,染成蓝绿色
暂无品牌问答