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Express® E. coli Lysis Reagent  |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

NEBExpress® E. coli Lysis Reagent is a chemical lysis solution composed of a proprietary mix of non-ionic and zwitterionic detergents and Tris-based buffer. It allows disruption of E. coli cells without denaturing soluble proteins.

Figure 1: Optimal protein extraction with NEBExpress T4 Lysozyme in the presence of NEBExpress E. coli Lysis Reagent.

Express® E. coli Lysis Reagent  |

T7 Express E. coli  expressing vGFP was lysed with NEBExpress T4 Lysozyme (T4L) in 50 mM Tris-HCl pH 7.5 or in NEBExpress E. coli Lysis Reagent (NEB #P8116S): 4 UOD600 of cell pellet were resuspended in 200 µL of Tris buffer or NEBExpress E. coli Lysis Reagent and lysed for 5 minutes at room temperature in the presence of NEBExpress T4 Lysozyme at 0, 0.1 or 1 ug per 1 mL of cell suspension. The soluble proteins were harvested by centrifugation and analyzed on SDS-PAGE.

产品类别:
Competent Cells Products

应用:
High-throughput cloning and automation solutions,
Protein Expression in E. Coli,
Protein Expression

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P8116S     25    
        NEBExpress® E. coli Lysis Reagent P8116SVIAL 25 1 x 100 ml 1 X
    • P8116L     25    
        NEBExpress® E. coli Lysis Reagent P8116SVIAL 25 5 x 100 ml 1 X

  • 优势和特性

    Features

    • Scalable lysis reactions from small to large bacterial cell pellets and compatible with high throughput workflows
    • Suitable for extracting proteins, especially thermosensitive proteins vulnerable to mechanical lysis procedures
    • Fast and simple protocol: harvested cells are resuspended and briefly incubated in the Lysis Reagent to extract soluble proteins. Insoluble cell debris is removed by centrifugation, and the clarified extract is ready to use and compatible with affinity resins.
    • Compatible with analyses such as SDS-PAGE, Western blots, activity assays, immunoprecipitation, and downstream purification 
    • Suitable for disrupting most Gram-negative bacterial cells
    • Small (100 ml) and large kits (5 x 100 ml) are suitable for lysing 20 to 100 grams of cells, respectively.
     

  • 相关产品

    相关产品

    • m0303-dnase-i-rnase-free
    • T7 表达 lysY E. coli 感受态细胞（高效级）
    • BL21(DE3) E. coli 感受态细胞
    • T7 表达 E. coli 感受态细胞（高效级）
    • Amylose Resin
    • Amylose Resin High Flow
    • 几丁质树脂
    • NEBExpress® Ni-NTA 磁珠
    • NEBExpress® Ni Spin Columns
    • NEBExpress® Ni Resin
    • E8201 NEBExpress MBP 融合表达及纯化系统
    • IMPACT™ Kit
    • 亲水性链霉亲和素磁珠
    • SNAP-捕获磁珠
    • Color Prestained Protein Standard, Broad Range (10-250 kDa)
    • 蓝色蛋白上样染料
    • NEBExpress® T4 Lysozyme
    • 微球菌核酸酶

  • 注意事项

    1. NEBExpress E. coli Lysis Reagent is compatible with: 

      • Tris, phosphate, and PIPPS-based buffers ranging from pH 5.5-10.
      • EDTA. To prevent interference with metal affinity resins use < 0.1-0.5 mM EDTA. If using NEBExpress Ni Resin (NEB #S1428S) or Ni Spin Columns (NEB #S1427S/L), there is no limitation on EDTA use, as the resin is chemically resistant to EDTA chelation.
      • < 1 M NaCl (precipitation will occur at or above 1 M NaCl).
      • Protease inhibitors 
      • Reducing agent such as DTT, β-mercaptoethanol, TCEP, THP. To prevent interference with metal affinity resins, use 1-2 mM β-mercaptoethanol or 1-5 mM TCEP. NEBExpress Ni Resin (NEB #S1428) and Ni Spin Columns (NEB #S1427S/L) are both chemically resistant to these reducing agents. 
      • BCA Protein assay
      • SDS-PAGE and Western blots
      • 1 µL NEBExpress® T4 Lysozyme (NEB# P8115) per 1 mL of lysate for improved protein extraction efficiency. The lysate may become viscous due to the release of DNA. For optimal lysis yield, add 1mM CaCl2 and 1 µL of Micrococcal Nuclease (NEB #M0247) per 1 mL of lysate, mix and incubate for 5 minutes at room temperature.
       
    2. NEBExpress E. coli Lysis Reagent can be added directly to frozen cell pellets; however, allow this mixture to reach room temperature as the lysis will be more efficient.
    3. Use 0.025 – 0.075 mL NEBExpress E. coli Lysis Reagent per unit OD600 (UOD600) cells harvested. For example, for a 5 mL culture harvested at OD600 of 1, 5 UOD600 of cells are obtained (5 mL x 1 OD600 = 5 UOD600), thus requiring 0.125 to 0.375 mL of Lysis Reagent. 
    4. If weighing the cell pellet, use 5 mL NEBExpress E. coli Lysis Reagent per 1 gram of cell paste. Note that cells resuspended in Lysis Reagent at higher cell densities will not lyse as efficiently.
    5. Extraction efficiency is strain dependent. Efficiency is highest with BL21 strains, its derivatives with DE3 or lysY, E. coli K-12 strains, and various other Gram-negative bacteria (e.g., Salmonella, Vibrio, Pseudomonas).
    6. Storing NEBExpress E. coli Lysis Reagent at 4°C or below may cause precipitation of the detergents. However, the lysis efficiency of the reagent can be fully restored by incubation at room temperature with gentle mixing to dissolve any precipitate.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for cell lysis using NEBExpress E. coli Lysis Reagent (NEB #P8116)

FAQs & 问题解决指南

  • FAQs

    1. How much NEBExpress® E. coli Lysis Reagent should I use?
    2. Why do I see precipitation in the NEBExpress® E. coli Lysis Reagent?
    3. Can I use NEBExpress® E. coli Lysis Reagent that has been frozen?
    4. Should I chill the NEBExpress® E. coli Lysis Reagent prior to use?
    5. Should I freeze the cell pellets prior to lysing with NEBExpress® E. coli Lysis Reagent?
    6. What buffer is present in the NEBExpress® E. coli Lysis Reagent?
    7. Can I supplement the NEBExpress® E. coli Lysis Reagent with NaCl?
    8. Can I change the pH of NEBExpress® E. coli Lysis Reagent?
    9. Is the NEBExpress® E. coli Lysis Reagent compatible with EDTA?
    10. Can I add a protease inhibitor to the NEBExpress® E. coli Lysis Reagent reaction?
    11. Do I need to add lysozyme to aid in lysis?
    12. My lysate is viscous; how can I resolve this?
    13. Can I use a reducing agent with NEBExpress® E. coli Lysis Reagent?
    14. Does NEBExpress® E. coli Lysis Reagent lyse gram-positive bacterial cells or yeast cells?
    15. Does NEBExpress® E. coli Lysis Reagent lyse gram-negative bacterial cells?
    16. My protein forms inclusion bodies. Can I still use this reagent to lyse my cells?
    17. Can I add NEBExpress® E. coli Lysis Reagent directly to my cell culture to lyse the cells?

  • 问题解决指南

    • Troubleshooting Guide for NEBExpress E. coli Lysis Reagent (NEB # P8116)

Express® T4 Lysozyme |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

NEBExpress® T4 Lysozyme is a recombinant murein hydrolase that breaks down the bacterial cell wall by hydrolyzing the β-1,4 linkage between N-acetylmuramic acid and N-acetylglucosamine in the peptidoglycan of prokaryotic cells (gram-negative and some gram-positive bacteria).  It can be used to extract soluble proteins, membrane proteins, DNA, RNA, or metabolites. 

Source: An E. coli strain that carries the bacteriophage T4 gene e.

Figure 1: NEBExpress T4 Lysozyme is 200-fold more active than chicken egg white lysozyme.

Express® T4 Lysozyme |

T7 Express E. coli  expressing vGFP was lysed with NEBExpress T4 Lysozyme (T4L) or Chicken egg white lysozyme (ChL): 4 UOD600 of cell pellet were resuspended in 200 µL of 50 mM Tris-HCl pH 7.5 and lysed for 5 minutes at room temperature in the presence of NEBExpress T4 Lysozyme or Chicken egg white lysozyme at 0.1, 1, 10, 100 or 200 ug enzyme per 1 mL of cell suspension. The soluble proteins were harvested by centrifugation and analyzed on SDS-PAGE. 

Figure 2: Optimal protein extraction with NEBExpress T4 Lysozyme in the presence of NEBExpress E. coli Lysis Reagent.

Express® T4 Lysozyme |

T7 Express E. coli  expressing vGFP was lysed with NEBExpress T4 Lysozyme (T4L) in 50 mM Tris-HCl pH 7.5 or in NEBExpress E. coli Lysis Reagent (NEB #P8116S): 4 UOD600 of cell pellet were resuspended in 200 µL of Tris buffer or NEBExpress E. coli Lysis Reagent and lysed for 5 minutes at room temperature in the presence of NEBExpress T4 Lysozyme at 0, 0.1 or 1 ug per 1 mL of cell suspension. The soluble proteins were harvested by centrifugation and analyzed on SDS-PAGE.

 
产品类别:
Competent Cells Products,
E. coli Protein Expression Strains Products,
Protein Expression Products

应用:
Protein Expression in E. Coli,
Protein Expression

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P8115S     -20    
        NEBExpress® T4 Lysozyme P8115SVIAL -20 1 x 200 µg 1 mg/ml
    • P8115L     -20    
        NEBExpress® T4 Lysozyme P8115SVIAL -20 5 x 200 µg 1 mg/ml

  • 特性和用法

    贮存溶液

    50 mM Tris-HCl
    100 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    0.03% Poloxamer 188
    50% Glycerol
    pH 7.5 @ 25°C

  • 优势和特性

    Features

    • NEBExpress T4 Lysozyme is 200-fold more active than chicken egg white lysozyme.
    • Lysis reactions are scalable and compatible with high throughput workflows.
    • Lysis efficiency increases 2-fold when used in combination with NEBExpress E. coli Lysis Reagent (NEB #P8116S).
    • Fast and non-mechanical bacterial lysis; the lysate is ready to use and compatible with affinity resins.
    • Suitable for lysing thermosensitive proteins vulnerable to mechanical lysis procedures
    • Compatible with downstream applications such as SDS-PAGE, Western blot, activity assays, immunoprecipitation, and downstream purification 
    • Small (200 µg) and large (5 x 200 µg) formats are suitable for lysing 7 or 35 grams of cells, respectively.
    • Recombinant and animal free.
     

  • 相关产品

    相关产品

    • P8116 NEBExpress E coli Lysis Reagent
    • 微球菌核酸酶
    • m0303-dnase-i-rnase-free
    • BL21(DE3) E. coli 感受态细胞
    • T7 表达 E. coli 感受态细胞（高效级）
    • Amylose Resin
    • 几丁质树脂
    • NEBExpress® Ni Spin Columns
    • NEBExpress® Ni Resin
    • Monarch® 基因组 DNA 提取试剂盒
    • T3060 Monarch HMW DNA Extraction Kit for Tissue

  • 注意事项

    1. NEBExpress T4 Lysozyme is compatible with: 

      • Tris, HEPES and sodium phosphate buffers ranging from pH 6.5 – 9.5; optimal pH is 7.5 – 8.5. 
      • 0.5 – 2 mM EDTA, which will improve lysis efficiency on gram negative bacteria. EDTA concentrations above 2mM start inhibiting enzyme activity, with a 50% loss at 5 mM EDTA.  
      • ≤ 25 mM NaCl; enzymatic activity is inhibited by 80% at 50 mM NaCl 
      • ≤ 1 mM MgCl2; enzymatic activity is inhibited by 80% at 5 mM MgCl2 
      • ≤ 2% Triton X-100 or Tween 20
      • Protease inhibitor cocktails (with ≤ 2 mM EDTA) 
      • Reducing agent such as DTT, β-mercaptoethanol, TCEP and THP. 
      • For optimal lysis performance, include NEBExpress E. coli Lysis Reagent (NEB #P8116S)
    2. NEBExpress T4 Lysozyme can be added directly to resuspended cells or pre-mixed with the buffer prior to resuspension of the cell pellet.
    3. Use 1 µL of NEBExpress T4 Lysozyme per 1 mL of cells resuspended at 10 UOD600. For example, for a 5 mL culture harvested at OD600 of 2, 10 UOD600 of cells are obtained (5 mL x 2 OD600 = 10 UOD600). 
    4. If weighing the cell pellet, use 30 µL of NEBExpress T4 Lysozyme per 1 gram of cell paste resuspended in 30 mL buffer. 
    5. The lysate may become viscous due to the release of DNA. For optimal lysis yield, eliminate viscosity with Micrococcal Nuclease (NEB #M0247).  Add 1mM CaCl2 and 1 uL of Micrococcal Nuclease per 1 mL of lysate, mix and incubate for 5 minutes at room temperature.

    6. Lysis efficiency can be strain dependent. For hard to lyse samples, use 5 to 15 µL of NEBExpress T4 Lysozyme per 1 mL of resuspended cells.

    7. Cell pellets subjected to one freeze-thaw cycle prior to NEBExpress T4 Lysozyme treatment tend to lyse better. 

  • 参考文献

    1. Jensen, H.B., Kleppe K. (1972). Effect of Ionic Strength, pH, Amines and Divalent Cations on the Lytic Activity of T4 Lysozym. Eur. J. Biochem. 28, 116-122.

操作说明、说明书 & 用法

  • 操作说明

    1. NEBExpress® T4 Lysozyme Lysis Protocol (NEB #P8115)
    2. Protocol for Genomic DNA Extraction from Gram-negative Bacteria using NEBExpress® T4 Lysozyme (NEB #P8115) and the Monarch® Genomic DNA Purification Kit (NEB #T3010)
    3. Protocol for Genomic DNA Extraction from Gram-positive Bacteria using NEBExpress® T4 Lysozyme (NEB #P8115) and the Monarch® Genomic DNA Purification Kit (NEB #T3010)

FAQs & 问题解决指南

  • FAQs

    1. How are NEBExpress® T4 Lysozyme and chicken egg white lysozyme different?
    2. Can I use NEBExpress® T4 Lysozyme for DNA or RNA extraction?
    3. What buffers are compatible with NEBExpress® T4 Lysozyme?
    4. Can reducing agents be used when lysing with NEBExpress® T4 Lysozyme?
    5. Is it ok to dilute NEBExpress® T4 Lysozyme before adding it to my resuspended cells?
    6. What are the optimal incubation temperature and time for lysing cells with NEBExpress® T4 Lysozyme?
    7. Is NEBExpress®T4 Lysozyme active on organisms other than E. coli?
    8. After incubation with NEBExpress® T4 Lysozyme, the lysate became viscous; how can I resolve this?
    9. Should cell pellets be frozen prior to lysing with NEBExpress® T4 Lysozyme?
    10. Can I use NEBExpress® T4 Lysozyme in a standard lysozyme activity assay using a fluorescent substrate (4-methylumbelliferone)?

  • 问题解决指南

    • Troubleshooting Guide for NEBExpress E. coli Lysis Reagent (NEB # P8116)

Express® Competent E. coli (High Efficiency) |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

重点

  • Enhanced BL21 derivative ideal for Plac, Ptac, Ptrc expression vectors
  • Deficient in proteases Lon and OmpT
  • Resistant to phage T1 (fhuA2)
  • Does not restrict methylated DNA (McrA–, McrBC–, EcoBr–m–, Mrr–)
  • Free of animal products

基因型

fhuA2 [lon] ompT gal sulA11 R(mcr-73::miniTn10–TetS)2 [dcm] R(zgb-210::Tn10–TetS) endA1 Δ(mcrC-mrr)114::IS10

产品类别:
Non-T7 Expression,
E. coli Protein Expression Strains Products,
Protein Expression Products,

E. coli Expression Strains Products

应用:
Non-T7 Expression,
Protein Expression in E. Coli,
Protein Expression

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • C2523H     -80    
        SOC Outgrowth Medium B9020SVIAL 4 1 x 25 ml Not Applicable
        pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
        NEBExpress® Competent E. coli (High Efficiency) C2523HVIAL -80 20 x 0.05 ml Not Applicable
    • C2523I     -80    
        SOC Outgrowth Medium B9020SVIAL 4 1 x 25 ml Not Applicable
        pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
        NEBExpress® Competent E. coli (High Efficiency) C2523IVIAL -80 6 x 0.2 ml Not Applicable

  • 特性和用法

    Antibiotic for Plasmid Selection

    用于质粒筛选的抗生素 使用浓度
    Ampicillin 100 µg/ml
    Carbenicillin 100 µg/ml
    Chloramphenicol 33 µg/ml
    Kanamycin 30 µg/ml
    Tetracycline 15 µg/ml
    Streptomycin 25 µg/ml

    货运单

    • Ships on dry ice

    抗生素耐药性

    • nit

  • 优势和特性

    Features

    • Versatile non-T7 expression strain
    • Protease deficient

    应用特性

    Chemically competent E. coli cells suitable for high efficiency transformation and protein expression. Recommended host strain for pMAL Protein Fusion and Purification System.

     

    DNA Effects on Transformation Efficiency and Colony Output: Express® Competent E. coli (High Efficiency) |
    The optimal amount of DNA to use in a transformation reaction is lower than commonly recognized. Using clean, supercoiled pUC19, the efficiency of transformation is highest in the 100 pg-1 ng range. However, the total colonies which can be obtained from a single transformation reaction increase up to about 100 ng.
    Effect of heat shock time on NEBExpress competent E. coli transformation efficiency: Express® Competent E. coli (High Efficiency) |  
    50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except heat shock time varied from 0 to 80 seconds.
    Effect of DNA incubation time on NEB Express competent E. coli transformation efficiency: Express® Competent E. coli (High Efficiency) |
    50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except DNA incubation time varied from 0 to 40 minutes.

  • 相关产品

    相关产品

    • c3037-nebexpress-iq-competent-e-coli-high-efficiency

    单独销售的组分

    • SOC Outgrowth 培养基

  • 注意事项

    1. 注意：本产品含有害物质 DMSO。处理前请先查看 MSDS。
    2. 贮存和处理：感受态细胞应在 -80℃ 下贮存。在 -20℃ 贮存会导致转化效率显著降低。当细胞温度高于 -80℃ 时，即使不融化，细胞也会失去效率。

操作说明、说明书 & 用法

  • 操作说明

    1. High Efficiency Transformation (C2523)
    2. 5 Minute Transformation (C2523)
    3. Expression Using NEB Express (C2523)
    4. High Efficiency Transformation Protocol (NEB #C2523 and #C3037)

  • 使用指南

    • Additional E. coli Strain Genotypes
    • Competent E. coli Genome Sequences Tool
    • Genetic Markers
    • Making Unmethylated (Dam- Dcm-) DNA
    • McrA, McrBC and EcoKI Strain Phenotypes
    • Restriction of Foreign DNA by E. coli K-12

  • 应用实例

    • Using the PURExpress® In Vitro Protein Synthesis Kit for Heterologous In Vitro Expression and Functional Screening of FMN-dependent Oxidoreductase Variants
    • Enhancing Transformation Efficiency

工具 & 资源

  • 选择指南

    • Characteristics of Select E.coli Strains
    • Competent Cell Selection Guide

FAQs & 问题解决指南

  • FAQs

    1. Does New England Biolabs offer strains of Competent E.coli suited for protein expression?
    2. How long should I incubate cells on ice after DNA has been added (NEB #C2523H and NEB #C2523I)?
    3. What are the solutions/recipes (C2523)?
    4. What are the strain properties (C2523)?
    5. What is the difference between NEB #C2523H and NEB #C2523I?
    6. Why are there no colonies or no growth in liquid culture (C2523)?
    7. Why is induced protein insoluble (C2523)?
    8. Why is there no protein visible on gel or no activity (C2523)?
    9. Can I store competent cells at -20°C instead of -80°C?
    10. Can the NEBExpress Competent E.coli (High Efficiency) be used for the expression of constructs containing a T7 promoter?
    11. What is the optimal heat shock time for this strain (NEB #C2523H and NEB #C2523I)?
    12. Which kind of transformation tubes should be used?
    13. What volume of DNA can be added into competent cells?
    14. What is the shelf life for this strain (NEB #C2523H and NEB #C2323I)?
    15. Are NEB’s competent cells compatible with the “Mix & Go” protocol?
    16. How should I store SOC Outgrowth Medium? The SOC I received with my competent cells recommends storage at either room temperature or 4°C, however, when I purchase it as a stand alone product, it recommends storing it at 4°C. Which is better?

Express® Iq Competent E. coli (High Efficiency) |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

重点

  • Enhanced BL21 derivative ideal for Plac, Ptac, Ptrc expression vectors
  • LacIq reduces basal expression
  • Ideal for pUC19 expression
  • Deficient in proteases Lon and OmpT
  • Resistant to phage T1 (fhuA2)
  • Does not restrict methylated DNA (McrA–, McrBC–, EcoBr–m–, Mrr–)
  • Better control of IPTG induced expression of non T7 plasmids
  • Free of animal products

基因型

MiniF lacIq (CamR) / fhuA2 [lon] ompT gal sulA11 R(mcr-73::miniTn10–TetS)2 [dcm] R(zgb-210::Tn10–TetS) endA1 Δ(mcrC-mrr)114::IS10

产品类别:
Non-T7 Expression,
E. coli Protein Expression Strains Products,
Protein Expression Products,

E. coli Expression Strains Products

应用:
Non-T7 Expression,
Protein Expression in E. Coli,
Protein Expression

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • C3037I     -80    
        NEBExpress® Iq Competent E. coli (High Efficiency) C3037IVIAL -80 6 x 0.2 ml Not Applicable

  • 特性和用法

    Antibiotic for Plasmid Selection

    用于质粒筛选的抗生素 使用浓度
    Ampicillin 100 µg/ml
    Carbenicillin 100 µg/ml
    Kanamycin 30 µg/ml
    Streptomycin 25 µg/ml
    Tetracycline 15 µg/ml

    货运单

    • Ships on dry ice

    抗生素耐药性

    • nit
    • cam

  • 优势和特性

    Features

    • Control of IPTG induced expression from Plac, Ptac,Ptrc and PT5-lacO
    • Protease deficient

    应用特性

    Chemically competent E. coli cells suitable for high efficiency transformation and protein expression, featuring control of IPTG induced expression with non-T7 plasmids.

    DNA Effects on Transformation Efficiency and Colony Output: Express® Iq Competent E. coli (High Efficiency) |
    The optimal amount of DNA to use in a transformation reaction is lower than commonly recognized. Using clean, supercoiled pUC19, the efficiency of transformation is highest in the 100 pg-1 ng range. However, the total colonies which can be obtained from a single transformation reaction increase up to about 100 ng.
    Effect of heat shock time on NEB Express Iq competent E. coli transformation efficiency: Express® Iq Competent E. coli (High Efficiency) |
    50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except heat shock time varied from 0 to 80 seconds.
    Effect of DNA incubation time on NEB Express Iq competent E. coli transformation efficiency: Express® Iq Competent E. coli (High Efficiency) |
    50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except DNA incubation time varied from 0 to 40 minutes.

  • 相关产品

    相关产品

    • c2523-neb-express-competent-e-coli-high-efficiency

    单独销售的组分

    • SOC Outgrowth 培养基

  • 注意事项

    1. 注意：本产品含有害物质 DMSO。处理前请先查看 MSDS。
    2. 贮存和处理：感受态细胞应在 -80℃ 下贮存。在 -20℃ 贮存会导致转化效率显著降低。当细胞温度高于 -80℃ 时，即使不融化，细胞也会失去效率。

操作说明、说明书 & 用法

  • 操作说明

    1. High Efficiency Transformation Protocol (C3037)
    2. 5 Minute Transformation Protocol (C3037)
    3. Protocol for Expression Using NEB Express Iq (C3037)
    4. High Efficiency Transformation Protocol (NEB #C2523 and #C3037)

  • 使用指南

    • Additional E. coli Strain Genotypes
    • Competent E. coli Genome Sequences Tool
    • Genetic Markers
    • McrA, McrBC and EcoKI Strain Phenotypes

  • 应用实例

    • AppNote_Enhancing_Transformation_Efficiency
    • Enhancing Transformation Efficiency

工具 & 资源

  • 选择指南

    • Characteristics of Select E.coli Strains
    • Competent Cell Selection Guide

FAQs & 问题解决指南

  • FAQs

    1. Does New England Biolabs offer strains of Competent E.coli suited for protein expression?
    2. How long should I incubate cells on ice after DNA has been added (NEB #C3037H and NEB #C3037I)?
    3. What are the solutions/recipes (C3037)?
    4. What are the strain properties (C3037)?
    5. What is the difference between NEB #C3037H and NEB #C3037I?
    6. What is the optimal heat shock time for this strain (NEB #C3037H and NEB #C3037I)?
    7. Why are there no colonies or no growth in liquid culture (C3037)?
    8. Why is induced protein insoluble (C3037)?
    9. Why is there no protein visible on gel or no activity (C3037)?
    10. Can I store competent cells at -20°C instead of -80°C?
    11. What is the shelf life for this strain (NEB #C3037H and NEB #C3037I)?
    12. Which kind of transformation tubes should be used?
    13. What volume of DNA can be added into competent cells?
    14. Are NEB’s competent cells compatible with the “Mix & Go” protocol?
    15. Is antibiotic selection necessary to maintain the MiniF-lacIq plasmid?