Highlights
- Boost Detection: Included BashingBeads ensure complete lysis of tough-to-lyse samples.
- Ultra-Pure: Ready for qPCR, Next-Gen Sequencing, arrays, etc.
- Simple: Fastest workflow (≤ 20 minutes).
Description
| Applicable For | All sensitive downstream applications such as qPCR and Next-Generation Sequencing. |
|---|---|
| Elution Volume | ≥ 35 µl |
| Equipment | Microcentrifuge, vortex, cell disruptor/pulverizer |
| Processing Time | ≤ 15 minutes |
| Processing Volume | ≤100 mg fungi or bacteria (wet weight), 109 bacterial cells, 108 yeast cells, or 107 mammalian cells |
| Purity | Typical A260/A280 & A260/A230 ≥ 1.8 |
| Sample Source | Fungal and bacterial cell cultures, spores, pollen, nematodes, as well as other microorganisms can also be sampled. |
| Size Range | Capable of recovering genomic DNA sized fragments from up to and above 40 kb. Typical fragment sizes range from 25 kb - 35 kb. If present, parasitic and viral DNA will also be recovered |
| Type | Total DNA |
| Yield | ≤ 25 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 optimizing 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: 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.
Q4: 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 | |
|---|---|---|---|---|
| D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
| D3004-2-50 | g-DNA Wash Buffer | 50 ml | $18.00 | |
| D3004-1-100 | Genomic Lysis Buffer | 100 ml | $60.00 | |
| D3004-5-15 | DNA Pre-Wash Buffer | 15 ml | $10.00 | |
| C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
| C1057-50 | Zymo-Spin III-F Filters | 50 Pack | $59.00 | |
| D6001-3-40 | BashingBead Buffer | 40 ml | $29.00 | |
| C1078-50 | Zymo-Spin IICR Columns | 50 Pack | $55.00 | |
| S6012-50 | ZR BashingBead Lysis Tubes (0.1 & 0.5 mm) | 50 Tubes | $101.00 |
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大家好,
我是今年刚毕业找到一份实验室研究工作的小猫,以前常作蛋白合成和分子克隆的,新实验室的老板想要研究些信号通路问题,具体就是研究一个病毒的致病途径是什么,定了大方向后我就扎到了文献的海洋中,可是越看就越有一个问题不明白.
就是我能找到的几乎所有文章,都是用的癌细胞做的模型,比如说hela,就是乳头瘤,293,就是肾瘤,还有SH5Y,就是神经瘤.我不太懂,就是病毒感染人的时候,不是感染的正常细胞吗,用病毒来感染癌细胞,然后在这些癌细胞中检测病毒的作用阿，信号通路之类的，要怎么绕过癌细胞其实和正常细胞不一样这个问题呢？
我看大部分文献都不在这方面作解释说明，仿佛都是约定俗成的用癌细胞，我对信号通路研究也是啥都不懂，两眼抹黑，求各路大神赐教ORZ
【2】信号：信号是数据的电磁编码或电子编码。和数据一样，信号也分为模拟信号和数字信号。模拟信号是指电信号的参量是连续取值的，其特点是幅度连续。常见的模拟信号有电话、传真和电视信号等。数字信号是离散的，从一个值到另一个值的改变是瞬时的，就像开启和关闭电源一样。数字信号的特点是幅度被限制在有限个数值之内。常见的数字信号有电报符号、数字数据等。信号是运载消息的工具，是消息的载体。从广义上讲，它包含光信号、声信号和电信号等。
全部的：www.cell.com/snapshots
信号通路1.zip(33848.99k)
信号通路2.zip(24706.36k)
细胞信号转导分析的生物信息学方法:
a)选择关键词，从GO数据库中寻找相关基因，比如extracellular表示为分泌蛋白
b)通过GO,BioCarta和Kegg信号通路数据分析给定基因所属的信号通路，功能分类等
c)比较多组基因按功能，通路分组在统计学上的差异，从而得到各组基因的功能差异
d)新信号通路的分析
回复：【资源】很有价值的信号通路分析平台方法!-丁香园论坛
请问哪种神经细胞可以代替原代海马神经元培养细胞？PC12?HT-22?SH-SY5Y?BV-2一般都适用于什么模型，大家一般用哪个呢
pathwaybuild
→MAPKK→MAPK→进入细胞核→其它激酶或基因调控蛋白（转录因子） 的磷酸化修钸。
信号通路的组成： 配体――生长因子； RTK—酪氨酸；
接头蛋白（生长因子受体接头蛋白-2， GRB-2）； GRF－－鸟苷酸释放因子； Ras—GTP 结合蛋白； Raf――是丝氨酸/苏氨酸（Ser/Thr）
蛋白激酶（称 MAPKKK）。
主要功能： 调节细胞的增殖与分化， 促进细胞存活， 以及细胞代谢过程中的调节与校正。
