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EZ-Tn5? Promoter Insertion Kit/EZ-Tn5? 啟動子插入試劑盒
| 貨號 | EZI03T7 | 售價（元） | 7312 |
| 規(guī)格 | 10 rxns | CAS號 |
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產(chǎn)品信息
|
貨號 |
產(chǎn)品名稱 |
規(guī)格 |
價格/元 |
|
TNP92110 |
EZ-Tn5 Transposase/EZ-Tn5轉(zhuǎn)座酶 |
10 units |
6418 |
|
EZI982K |
EZ-Tn5? <KAN-2> Insertion Kit/EZ-Tn5? <KAN-2>插入試劑盒 |
10 rxns |
6950 |
|
EZI921T |
EZ-Tn5? <TET-1> Insertion Kit EZ-Tn5? <TET-1>插入試劑盒 |
10 rxns |
6999 |
|
EZI912D |
EZ-Tn5? <DHFR-1> Insertion Kit/EZ-Tn5? <DHFR-1>插入試劑盒 |
10 rxns |
7060 |
|
EZI03T7 |
EZ-Tn5? <T7/KAN-2> Promoter Insertion Kit/EZ-Tn5? <T7/KAN-2>啟動子插入試劑盒 |
10 rxns |
7312 |
|
EZI011RK |
EZ-Tn5? <R6Kγori/KAN-2> Insertion Kit/EZ-Tn5? <R6Kγori/KAN-2>插入試劑盒 |
10 rxns |
7517 |
|
CIS40025 |
CopyCutter? Induction Solution/CopyCutter?誘導(dǎo)試劑 |
25 mL |
856 |
Transposons are mobile DNA sequences found in the genomes of prokaryotes and eukaryotes. Transposon tagging has long been recognized as a powerful research tool for randomly distributing primer binding sites, creating gene“knockouts”, and introducing a physical tag or a genetic tag into large target DNAs. One frequently used transposition system is the Tn5 system isolated from gram-negative bacteria. Though a naturally occurring transposition system, the Tn5 system can be readily adapted for routine use in research laboratories for the following reasons:
1) Tn5 transposase is a small, single subunit enzyme that has been cloned and purified to high specific activity.
2) Tn5 transposase carries out transposition without the need for host cell factors.
3) Tn5 transposon insertions into target DNA are highly random.
4) Tn5 transposition proceeds by a simple“cut and paste”process. Although the chemistry is unique, the result is similar to using a restriction endonuclease, with random sequence specifi city, accompanied by a DNA ligase activity.
5) Tn5 transposase will transpose any DNA sequence contained between its short 9 basepair Mo saic End (ME) Tn5 transposase recognition sequences.
In 1998 Goryshin and Reznikoff1demonstrated that a fully functional Tn5 transposition system could be reconstituted in vitro. Additionally, the transposition efficiency of this system has been increased more than 1,000-fold compared to wild-type Tn5 by introducing mutations in the transposase gene and in the 19-bp Tn5 ME transposase recognition sequence.
Lucigen's EZ-Tn5 Transposon Tools (kits and reagents) are based on the hyperactive Tn5 transposition system developed by Goryshin and Reznikoff.
轉(zhuǎn)座子是在原核生物和真核生物的基因組中發(fā)現(xiàn)的可移動DNA序列。 長期以來，轉(zhuǎn)座子標(biāo)記一直被認為是一種強大的研究工具，可用于隨機分布引物結(jié)合位點，創(chuàng)建基因“敲除”并將物理標(biāo)簽或遺傳標(biāo)簽引入大靶DNA。 一種常用的轉(zhuǎn)座系統(tǒng)是從革蘭氏陰性細菌中分離的Tn5系統(tǒng)。 盡管是自然發(fā)生的換位系統(tǒng)，但由于以下原因，Tn5系統(tǒng)可以很容易地用于研究實驗室的常規(guī)使用：
1）Tn5轉(zhuǎn)座酶是一種小的單亞基酶，已被克隆并純化成高比活性。
2）Tn5轉(zhuǎn)座酶無需宿主細胞因子即可進行轉(zhuǎn)座。
3）Tn5轉(zhuǎn)座子插入目標(biāo)DNA的隨機性很高。
4）Tn5轉(zhuǎn)座通過簡單的“剪切和粘貼”過程進行。 盡管化學(xué)性質(zhì)獨特，但結(jié)果類似于使用限制性核酸內(nèi)切酶，具有隨機序列特異性，并伴有DNA連接酶活性。
5）Tn5轉(zhuǎn)座酶將轉(zhuǎn)置其短9個堿基對的Mo saic End（ME）Tn5轉(zhuǎn)座酶識別序列之間的任何DNA序列。
1998年，Goryshin和Reznikoff1證明可以在體外重建功能齊全的Tn5轉(zhuǎn)座系統(tǒng)。 此外，通過在轉(zhuǎn)座酶基因和19 bp Tn5 ME轉(zhuǎn)座酶識別序列中引入突變，與野生型Tn5相比，該系統(tǒng)的轉(zhuǎn)座效率已提高了1,000倍以上。
Lucigen的EZ-Tn5轉(zhuǎn)座子工具（試劑盒和試劑）基于Goryshin和Reznikoff開發(fā)的高活性Tn5轉(zhuǎn)座系統(tǒng)。
優(yōu)點：
Insert a kanamycin, tetracycline, or DHFR selectable marker into any DNA sequence in vitro
Skip primer walking - simplify Sanger sequencing of large DNA inserts
Speed functional analysis without subcloning - create libraries of random mutants from purified DNA
Minimize insertion bias with the hyperactive Tn5 system, known for highest level of randomness
將卡那霉素，四環(huán)素或DHFR選擇標(biāo)記插入體外任何DNA序列
跳過引物行走-簡化大型DNA插入片段的Sanger測序
無需亞克隆即可加速功能分析-從純化的DNA創(chuàng)建隨機突變體庫
使用高度活躍的Tn5系統(tǒng)將插入偏差降至最低
組成成分：
EZ-Tn5? Transposase 10 U ：儲存在-20℃
EZ-Tn5? <R6Kγori/KAN-2> Transposon：儲存在-20℃
EZ-Tn5? 10X Reaction Buffer：儲存在-20℃
EZ-Tn5? 10X Stop Solution:儲存在-20℃
KAN-2 FP-1 Forward Primer:儲存在-20℃
R6KAN-2 RP-1 Reverse Primer:儲存在-20℃
pUC19/3.4 Control Target DNA:儲存在-20℃
Sterile Water:儲存在-20℃
數(shù)據(jù)：
產(chǎn)品優(yōu)點- Insert a kanamycin, tetracycline, or DHFR selectable marker into any DNA sequence in vitro
- Skip primer walking - simplify Sanger sequencing of large DNA inserts
- Speed functional analysis without subcloning - create libraries of random mutants from purified DNA
- Minimize insertion bias with the hyperactive Tn5 system, known for highest level of randomness
- 將卡那霉素，四環(huán)素或DHFR選擇標(biāo)記插入體外任何DNA序列
- 跳過引物行走-簡化大型DNA插入片段的Sanger測序
- 無需亞克隆即可加速功能分析-從純化的DNA創(chuàng)建隨機突變體庫
- 使用高度活躍的Tn5系統(tǒng)將插入偏差降至最低
產(chǎn)品應(yīng)用- Introducing a phage T7 RNA polymerase transcriptional promoter into any DNA.
- Faster sequencing of large DNA molecules, as compared to primer walking, random subcloning, or generating nested deletions with exonuclease III and mung bean nuclease.
- Making insertion mutants or gene “knockouts” in vitro.
- Introducing a kanamycin resistance selection marker into any DNA