![SMOBIO/[TP2000] ExcelTaq™ Blood Direct DNA Polymerase, (5 U/μl, 500 U)/5 U/μl, 500 U/TP2000](images/SMOBIO/image.jpg)
Description
The ExcelTaq™ Blood Direct DNA Polymerase is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct DNA Polymerase is highly tolerant in the presence of PCR interfering/ inhibiting substances in blood, such as IgG, hemoglobin, and lactoferrin. ExcelTaq™ Blood Direct DNA Polymerase is compatible with most anticoagulants, such as citrate, EDTA, and heparin (Fig. 1). The ExcelTaq™ Blood Direct DNA Polymerase includes a pair of Positive Control Primers (CCR5) that are compatible with primate blood samples.
Features
5"→3" DNA polymerase activity
No detectable 3"→5" exonuclease (proofreading) activity
Generates PCR products with 3"-dA overhangs
Compatible with most anticoagulants
Applications
Direct amplification of DNA from blood samples
High throughput screening without DNA purification
Suitable for multiplex PCR
Storage
-20°C for 24 months
Compatible with most anticoagulants
ExcelTaq™ Blood Direct DNA Polymerase amplified 200 bp from differently treated blood using CCR5 specific primers. (M: DM2100)
Contents
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Storage Buffer
20 mM Tris-HCl pH 8.0, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerol
CCR5 control primer sequence
CCR5-F:5"-CTCCCAGGAATCATCTTTACC-3"
CCR5-R:5"-TCATTTCGACACCGAAGCAG-3"
Storage
-20°C for 24 months
Manual
Manual_TP2000_ExcelTaq™ Blood Direct DNA Polymerase
SDS
SDS_TP2000
Recommended PCR Condition
Blood | 3μl |
Forward primer | 0.1– 0.5 µM |
Reverse primer | 0.1– 0.5 µM |
5X Blood Direct Buffer | 10μl |
dNTPs | 0.2mM (each) |
Blood Direct DNA Polymerase | 0.25μl (1.25 U) |
H2O | to50 μl |
Total volume | 50µl |
Note: Mix well before PCR Reaction
Recommended PCRProgram
Steps | Temp. | Time | Cycles |
Templatedenature | 94°C | 3 min | 1 |
Denature | 94°C | 30 sec | 25-40 |
Annealing | 50-68°C* | 30 sec | |
Extension | 72°C | 60 sec/kb | |
Final extension | 72°C | 3 min | 1 |
*Optimal PCR condition variesaccording to primers’ thermodynamic properties.
Post amplificationanalysis
Centrifuge PCR reaction mixture at 1000 × g for 1 ~ 3 minutes. Load only the clear supernatant of the PCRproducts for gel electrophoresis analysis.
[TP2100] ExcelTaq™ 5X Blood Direct PCR Master Mix Kit
5"→3" DNA polymerase activity
None detectable 3"→5" exonuclease (proofreading) activity
Generates PCR products with 3"-dA overhangs
High throughput PCR
Execute PCR directly from blood samples
High reproducibility, less pipetting errors
Compatible with most anticoagulants
[TP5000] ExcelTaq™ Hot Start II DNA Polymerase
Aptamer-based hot start PCR
Reversible enzyme inactivation
Omits extra enzyme activation step
Convenient for room temperature PCR set-up
High yield and specificity of target amplicons
Wide range of amplicon length (up to 10 kb)
High sensitivity (as low as 1 fg of plasmid)
Aptamer-based hot start PCR
Reversible enzyme inactivation
Omits extra enzyme activation step
Convenient for room temperature PCR set-up
High yield and specificity of target amplicons
Wide range of amplicon length (up to 10 kb)
High sensitivity (as low as 1 fg of plasmid)
[TQ1200] ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX)
High Stability
Fast Hot Start
High Sensitivity
Low Background / High Specificity
Suitable for Fast Program
Smart Blue Contrast Dye
[TP1200] ExcelTaq™ 5X PCR Master Dye Mix
5’→3’ DNA polymerase activity
No detectable 3"→5" exonuclease (proofreading) activity
Generates PCR products with 3"-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
Includes tracking dye for direct loading after PCR
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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。
