SMOBIO/[CK1000] Champion™ E. coli Transformation Kit, 200 Rxn/s)"}]]> </ul> <a class=b,SMOBIO,,SMOBIO,+,,Array.Array.Array蚂蚁淘厂家直采.

**Prepare Competent Cells with High Transformation Efficiency Using Champion™ E.coli Transformation Kit**

**Prepare Competent Cells Directly from E.coli Colonies Using Champion™ E.coli Transformation Kit**

Time-saving Transformation Protocol

Kit Contents

Can Champion™ E. coli Transformation Kit be used to make competent cell of other bacteria?

What E.coli. strain is compatible to Champion™ E. coli Transformation Kit?

Will transformation efficiency be reduced when Champion™ competent cells are thawed, dispensed and refrozen repeatedly?

What is the advantage of thawing competent cells with circulating water instead of still water?

Is there any difference of transformation efficiency between using plating beads, bending glass rod and plating loop?

Is 1 second vortex critical for mixture the DNA?

Will heat shock affect the transformation efficiency?

Is it necessary to change the transformation procedure for transforming a large plasmid?

How to reduce the interference of the satellite colonies?

Culture Condition

E. coli Strains

Storage Condition

Thawing methods

Size of plasmid

Heat shock treatment

Plating methods

Concentration of antibiotic

High Fidelity PCR amplification

产品名称：SMOBIO/[CK1000] Champion™ E. coli Transformation Kit, 200 Rxn/s)"}]]>

产品编号：CK1000

市场价：¥0.00

场地：美国(厂家直采)

产品分类：其它>配件>影像显示>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：待定

品牌： SMOBIO

公司：SMOBIO

公司分类：

商品介绍

Description

Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 10⁸–10⁹cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps.

Features

Flexible– fresh culture, overnight culture, 4°C stored liquid culture or even colonies on agar plate can be used for transformation.
Fast and Easy– only few steps for preparation; suitable for time-saving transformation
High efficiency– up to 109 cfu/μg
Personalization– suitable for most E. coli strains

Kit Contents

Storage

4°C for 12 months

Component

Volume

Champion™ CC Buffer

20 ml

SMO-Broth™

2 x 100 ml

pUC19 Control Plasmid (10^-4 μg/μl)

5 µl

Instruction Manual

Champion™ Competent CellPreparation Card

Storage

4°C for 12 months

Manual

Manual_CK1000_Champion™ E. coli Transformation Kit

SDS

SDS_CK1000

Protocol card

Champion™ E. coli Transformation Kit are specially designed for making E.coli competent cells. Therefore, Champion™ E. coli Transformation Kit is not recommend to used for make other bacterial competent cells.

Most E.coli strains are compatible to Champion™ E. coli Transformation Kit. However, different E. coli strains vary in their ability to be transformed with DNA.

Efficiency of strains and their derivatives is listed below, when prepared with the Champion™ E. coli Transformation Kit.

E. coli strains

Efficiency (cfu/μg)

JM109

~5x108

XL-1 blue

~3x108

DH5α

~5x108

stbl 3

~2x108

BL21

~1x107

Rosetta 2

~1x107

*E. coli strains are liquid cultured following standard protocol for preparation of competent cells (at 25°C until OD600 reached ~0.5).

When Champion™ competent cells are thawed dispensed in aliquots and refrozen within 1 min, the transformation efficiency will be 10~20% reduced compared to the first time use.

Using circulating water can reduce the thawing time. Less thawing time shows higher efficiency than long thawing time.

The bending glass rod shows best efficiency and the plating loop show less efficiency than other method. For handle a lot samples at the same time the plating beads are recommended.

One second vortex provides more reliable transformation efficiency (1.1 times compare with mixed by finger tapping)

Heat shock treatment will enhance the efficiency about 1~2X versus non-heat shock method.

For large plasmid (> 6 kb), the heat shock method will significantly improves the efficiency and the recovery procedure also significantly improves the efficiency.

Using proper antibiotics of suitable concentration or using fresh antibiotics.

E. coli cells present high competence when freshly cultured from 1/100~1/20 dilution of overnight culture and incubated at 16~25°C prior to preparation. Higher temperature (37°C) leads to competence decrease.

Temperature	OD₆₀₀	Culturetime	Efficiency(cfu/μg)
16°C	0.4~0.8	12~20hr	~1x10⁹
25°C	0.4~0.8	3~8hr	~5x10⁸
37°C	0.4~0.8	1~1.5hr	~1x10⁸
25°C	1~2.5	4~10hr	~3x10⁸

E. coli cells in liquid culture are higher competent than colonies on plate.

Culture Condition

Efficiency (cfu/μg)

Liquidculture (fresh or overnight culture)

10⁸~10⁹

Freshcolonies (grown overnight at 37°C)

~2x10⁶

Oldcolonies (4°C storedfor 7 days)

~5x10⁵

Different E. coli strains vary in their ability to be transformed with DNA. Efficiency of strains and their derivatives is listed below, when prepared with the Champion™ E. coli Transformation Kit.

E. coli strains

Efficiency (cfu/μg)

JM109

~5x10⁸

XL-1 blue

~3x10⁸

DH5α

~5x10⁸

stbl 3

~2x10⁸

BL21

~1x10⁷

Rosetta 2

~1x10⁷

*E. coli strains are liquid cultured following standard protocol for preparation of competent cells (at 25°C until OD600 reached ~0.5).

To maintain high competence, E. coli should be stored at -70°C. Higher temperature (above -50°C) leads to competence decrease.

Shorter thawing time is more efficient than a longer thawing time. Slow thawing caused by power shortages and unstable freezers will result in decreased efficiency.

Plasmid size affect the efficiency greatly. The efficiency of transforming a supercoiled 2.7 kb is approximately 100 times higher than that of a 10 Kb plasmid (using time-saving transformation protocol). For large plasmids (> 6 kb), standard transformation protocol is recommended.

Heat shock treatment will enhance the efficiency about 1~2 folds versus non-heat shock method.

Bent glass rods show the greatest efficiency, while plating loop shows less efficiency than plating beads. When handling a large quantity of samples at the same time, plating beads are recommended.

Antibiotic concentration is critical to use of the time-saving transformation protocol.

Antibiotic

Concentration

Ampicillin (Ap)

20 μg/ml

Kanamycin (Km)

25 μg/ml

Tetracycline (Tc)

7.5 μg/ml

Chloramphenicol (Cm)

20 μg/ml

For plasmid size <6 Kb, the efficiency of kanamycin selection is usually 3~10 times less than the ampicillin selection. For plasmid size > 6 Kb, the efficiency of kanamycin selection is much lower than ampicillin. We suggest using the standard transformation protocol (with heat shock and recovery steps) to enhance the efficiency.

Champion™ CC Buffer

SMO-Broth™

pUC19 Control Plasmid (10^-4 μg/μl)

Instruction Manual

Champion™ Competent CellPreparation Card

Amplification of target gene with HiFi™ DNA polymerase to minimize error rate.

[TF1000] SMO-HiFi™ DNA Polymerase, (1 U/μl, 100 U)
[TF3000] G-HiFi™ DNA Polymerase, (1 U/μl, 100 U)

Gel electrophoresis

Staining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency.

Safe fluorescent dyes

[NS1000] FluoroVue™ Nucleic Acid Gel Stain (10,000X), 500 μl
[DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl
[DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 ml

Blue-light illuminator

[VE0100] B-BOX™ Blue Light LED Epi-illuminator

Ligation

Blund-end PCR amplicons can directly ligate with PCR cloning vector.

[CV1000] GetClone™ PCR Cloning Vector, 20 Rxn
[CV1100] GetClone™ PCR Cloning Vector II, 20 Rxn

Colony PCR

Analyze colonies with PCR master mix to save preparation time.

[TP1100] ExcelTaq™ 5X PCR Master Mix, 200 Rxn
[TP1200] ExcelTaq™ 5X PCR Master Dye Mix, 200 Rxn

品牌介绍

SMOBIO

SMOBIO成立于2004年，公司在开发和制造分子生物学试剂和测试工具方面表现优异。其分子生物学试剂是市场上最受欢迎的一部分。提供的蛋白质标记物包含具有广谱预染蛋白的即用型稳定混合物，范围从50bp到25kb。此外，SMOBIO提供各种DNA聚合酶和逆转录酶，可以针对不同的实验室应用进行优化。专注领域：蛋白质标记、DNA梯度、PCR相关产品、克隆载体、荧光加载和染色染料、RT-PCR相关产品、Taq DNA聚合酶、Western Marker、dNTP等。

2004年，SMOBIO Technology，Inc.迅速出现在生物技术行业。SMOBIO Technology，Inc.首先采用独特的发酵技术，旨在生产高质量的功能性食品成分。建立成功的分销网络后，我们将继续专注于高质量产品，并开发了最先进的生物分子测试工具。我们的工具旨在通过提供高质量的DNA阶梯和蛋白质标记物来促进分子生物学和蛋白质科学研究。这些创新产品已受到业界和学术界人士的高度评价。自2008年以来，SMOBIO Technology，Inc.已展示出非常可观的市场份额。客户满意的证据表明，SMOBIO Technology，Inc.竭尽全力生产高质量的产品。

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