W3110 Electro-competent cells电转感受态细胞 BioVector NTCC质粒载体菌种细胞基因保藏中心
- 价 格：¥19752
- 货 号：W3110 Electro-competent cells电转感受态细胞
- 产 地：北京
- BioVector NTCC典型培养物保藏中心
- 联系人：Dr.Xu, Biovector NTCC Inc.
电话：400-800-2947 工作微信:1843439339 (QQ同号)
邮件：Biovector@163.com
手机：18901268599
地址：北京
- 已注册
W3110 Electro-competent cells电转感受态细胞 BioVector NTCC质粒载体菌种细胞基因保藏中心 NTCC-W3110ECC W3110 Electro-competent cells 10x200uL.
Resistance:nalidixic acid. Storage:-80℃
Description
Classification Enterobacteriaceae, Escherichia
Strain Designations W3110
Application transformation host
Biosafety Level 1
Genotype F- mcrA mcrB IN(rrnD-rrnE)1 lambda-
Comments derived from strain K-12
A prototrophic derivative of K-12 (nearly wild type).
Protocols
Transformation into a plasmid vector
1) Thaw E. coli Electro-Cells (50 μl) in an ice bath just before use.
2) Add 1 - 2 μl of DNA solution into the thawed cell suspension.
3) Transfer the mixture of cells and DNA to a cold 0.1 cm electroporation cuvette.
4) After applying pulse, immediately add 1 ml of SOC medium (precooled in an ice bath).
5) Incubate by shaking (160 - 250 rpm) for 1 hour at 37℃.
6) Plate on selective media. Less than 100 μl is applied to a φ9 cm plate.
7) Incubate overnight at 37℃.
＊1 When the sample DNA solution contains salt(s), dilute it with TE buffer or distilled sterilized water. Alternatively, desalination by ethanol precipitation is recommended. Ethanol precipitation should be performed when the sample DNA is prepared for use with the DNA Ligation Kits.
＊2 We recommand using BIO-RAD MicroPulser and the electrical condition is 1.5 kV.
In the case of BIO-RAD Gene Pulser, standard electrical conditions are 200 Ω, 25 μF and 1.5 kV.
Please read before Proceeding
1) Place a tube of Electro-Cells in a dry ice / EtOH bath immediately upon removal from the -80℃ freezer. Keep the cells in the bath until you are ready to proceed.
2) For 50 μl of Electro-Cells, use no more than 10 ng of high purity DNA, or the transformation efficiency may be reduced.
3) If using large size of DNA( > 7 kb), transformation efficiency might decrease.
4) If you change the quantity of Electro-Cells electroporated, it may be necessary to reevaluate the conditions.
5) L-broth or φ b-broth can be used instead of SOC medium, but efficiency may be reduced.
・L-broth： 10 g Bacto tryptone, 5 g Bacto yeast extract, 5 g NaCl/1 L water, pH to 7.5 with 1 N NaOH, and autoclave.
・ψ b-broth： 5 g Bacto yeast extract, 20 g Bacto tryptone, 5 g MgSO4･7 H2O/1 L water, pH to 7.5 with 1 N KOH, and autoclave.
6) When diluting, use the medium which has been added in the step 4) .
7) When using X-Gal:
・ Add 20 mg/ml X-Gal (dissolved in dimethylformamide) at a ratio of 200 - 300 μl/100 ml agar media.
8) Once the Electro-Cells have been thawed, refreezing for storage is not recommended. If this is unavoidable, flash freeze the cells on dry ice/ethanol and store them promptly at -80℃. However, the transformation efficiency will be lowered by at least one order of magnitude.
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
TEL:+86-010-53513060
网址http://www.biovector.net [Supplier来源] http://www.biovector.net
Resistance:nalidixic acid. Storage:-80℃
Description
Classification Enterobacteriaceae, Escherichia
Strain Designations W3110
Application transformation host
Biosafety Level 1
Genotype F- mcrA mcrB IN(rrnD-rrnE)1 lambda-
Comments derived from strain K-12
A prototrophic derivative of K-12 (nearly wild type).
Protocols
Transformation into a plasmid vector
1) Thaw E. coli Electro-Cells (50 μl) in an ice bath just before use.
2) Add 1 - 2 μl of DNA solution into the thawed cell suspension.
3) Transfer the mixture of cells and DNA to a cold 0.1 cm electroporation cuvette.
4) After applying pulse, immediately add 1 ml of SOC medium (precooled in an ice bath).
5) Incubate by shaking (160 - 250 rpm) for 1 hour at 37℃.
6) Plate on selective media. Less than 100 μl is applied to a φ9 cm plate.
7) Incubate overnight at 37℃.
＊1 When the sample DNA solution contains salt(s), dilute it with TE buffer or distilled sterilized water. Alternatively, desalination by ethanol precipitation is recommended. Ethanol precipitation should be performed when the sample DNA is prepared for use with the DNA Ligation Kits.
＊2 We recommand using BIO-RAD MicroPulser and the electrical condition is 1.5 kV.
In the case of BIO-RAD Gene Pulser, standard electrical conditions are 200 Ω, 25 μF and 1.5 kV.
Please read before Proceeding
1) Place a tube of Electro-Cells in a dry ice / EtOH bath immediately upon removal from the -80℃ freezer. Keep the cells in the bath until you are ready to proceed.
2) For 50 μl of Electro-Cells, use no more than 10 ng of high purity DNA, or the transformation efficiency may be reduced.
3) If using large size of DNA( > 7 kb), transformation efficiency might decrease.
4) If you change the quantity of Electro-Cells electroporated, it may be necessary to reevaluate the conditions.
5) L-broth or φ b-broth can be used instead of SOC medium, but efficiency may be reduced.
・L-broth： 10 g Bacto tryptone, 5 g Bacto yeast extract, 5 g NaCl/1 L water, pH to 7.5 with 1 N NaOH, and autoclave.
・ψ b-broth： 5 g Bacto yeast extract, 20 g Bacto tryptone, 5 g MgSO4･7 H2O/1 L water, pH to 7.5 with 1 N KOH, and autoclave.
6) When diluting, use the medium which has been added in the step 4) .
7) When using X-Gal:
・ Add 20 mg/ml X-Gal (dissolved in dimethylformamide) at a ratio of 200 - 300 μl/100 ml agar media.
8) Once the Electro-Cells have been thawed, refreezing for storage is not recommended. If this is unavoidable, flash freeze the cells on dry ice/ethanol and store them promptly at -80℃. However, the transformation efficiency will be lowered by at least one order of magnitude.
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
TEL:+86-010-53513060
网址http://www.biovector.net [Supplier来源] http://www.biovector.net
- 公告/新闻
