NEST BioFactory

ig 10B
Intact Genomics ig 10B chemically competent cells (E. coli) are suitable for high efficiency transformation in a wide variety of applications such as cloning and sub-cloning. This derivative of DH10B provides the highest efficiency in the industry.

• Most efficient chemically competent cells in the industry (up to 10X more efficient!)
• May easily be used in place of NEB 10-Beta, TOP10 from Invitrogen or other common brands
• Significantly more affordable

Specifications
Competent cell type:                Chemically competent
Derivative of:                            DH10B
Species:                                   E. coli
Format:                                    Tubes
Transformation efficiency:         â¥1.0 x 1010 cfu/µg pUC19 DNA
Blue/white screening:               Yes
Shipping condition:                   Dry ice

Quality Control
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Reagents Needed for One Reaction
ig 10B chemically competent cells:     50 µl
DNA (or pUC19 Control, 10 pg/µl):          1 µl
Recovery medium:                                  1 ml

Contents & Storage

1. ig 10B competent cells: -80 °C
2. pUC19 control DNA: -20 °C
3. Recovery medium: 4 °C

Genomic Features
ig 10B (DH10B derivative) chemically competent cells have the following features:

• 80lacZM15 marker provides α-complementation of the β-galactosidase gene with blue/white screening
• mcrA genotypic marker and the mcrBC, mrr deletion allows for cloning DNA that contains methylcytosine and methyladenine

Genotype
F â mcrA (mrr-hsdRMS-mcrBC) endA1 recA1 80dlacZM15 lacX74 araD139 (ara, leu)7697 galU galK rpsL (StrR) nupG -

General Guidelines
Follow these guidelines when using ig 10B chemically competent E. coli.

• Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical lysis caused by pipetting.
• Thaw competent cells on ice, and transform cells immediately following thawing. After adding DNA, mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.

Protocol

• Remove competent cells from the -80 °C freezer and thaw completely on wet ice (10-15 minutes).
• Aliquot 1-5 µl (1 pg-100 ng) of DNA to the chilled microcentrifuge tubes on ice.
• When the cells are thawed, add 50 µl of cells to each DNA tube on ice and mix gently by tapping 4-5 times. For the pUC19 control, add 1 µl of (10 pg/µl) DNA to a chilled microcentrifuge tube, prior to adding 50 µl of cells. Mix well by tapping.
• Incubate the cells with DNA on ice for 30 minutes.
• After 30 minute ice incubation, heat shock the cells at 42 °C for 45 seconds.
• Transfer the tubes to ice for 2 minutes.
• Add 950 µl of Recovery Medium or any other medium of choice to each tube.
• Incubate tubes at 37 °C for 1 hour at 210 rpm.
• Spread 50 µl to 200 µl from each transformation on Pre-warmed selection plates.
• Incubate the plates overnight at 37 °C.