igMax DH10B ElectroCompetent Cells

Description

Intact Genomics igMax^TM DH10B derivative ElectroCompetent cells are suitable for demanding cloning situations, such as synthetic bio-applications, BAC cloning, assembling large and multi-DNA fragments, and large construct transformation or cloning difficult targets, requiring the greatest number of transformants possible. Utilizing proprietary manufacturing methods, these cells allow for effective transformation of all large DNA molecules (≥10kb up to 350kb!).

Intact Genomics igMax^TM DH10B cells vs. Competitors DH10B Derivative transformation efficiency

Extremely large DNA molecules can effectively be transformed at a rate much higher than ANY competitor, as shown in the figure above.

Specifications
Competent cell type:            ElectroCompetent
Derivative of:                         DH10B
Species:                                 E. coli
Format:                                  Tubes
Transformation efficiency:     ≥ 5 x 10¹⁰ cfu/µg pUC19 DNA
Blue/white screening:             Yes
Shipping condition:                 Dry ice

Reagents Needed for One Reaction
igMax^TM DH10B ElectroCompetent cells:  25 µl
DNA (or pUC19 Control, 10 pg/µl):    1 µl
Recovery medium:    1 ml

Storage
igMax^TM DH10B ElectroCompetent cells:-80 ºC
pUC19 control DNA: -20 ºC
Recovery medium:    4 ºC

Genomic Features
Intact Genomics igMax^TM ElectroCompetent DH10B cells have the following features:

• ≥5 x 10¹⁰cfu/µg efficiency with electroporation.
• 5~10×10⁷ for 100~150 kb large DNA

Genotype

F – mcrA ∆(mrr-hsdRMS-mcrBC) endA1 recA1 φ80dlacZ∆M15 ∆lacX74 araD139 ∆(ara, leu)7697 galU galK rpsL (StrR) nupG λ-

Quality Control
Transformation efficiency is tested by using pUC19, ~50kb, and >100kb plasmids. The pUC19 control DNA is supplied with the kit and using the protocol given below. Transformation efficiency should be ≥5 x 10¹⁰ CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines
Follow these guidelines when using Intact Genomics igMax^TM  DH10B ElectroCompetent cells:

• 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.

Note: A high-voltage electroporation apparatus such as Bio-Rad Gene Pulser II #165-2105, capable of generating field strengths of 16 kV/cm is required.

Calculation of Transformation Efficiency
Transformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1µg of plasmid into a given volume of competent cells.
TE = Colonies/µg/Dilution
Transform 1 µl of (10 pg/µl) pUC19 control plasmid into 25 µl of cells, add 975 µl of Recovery Medium. Dilute 10 µl of this in 990 µl of Recovery Medium and plate 50 µl. Count the colonies on the plate the next day. If you count 250 colonies, the TE is calculated as follows:
Colonies = 250
µg of DNA = 0.00001
Dilution = 25/1000 x 10/1000 = 0.00025
TE = 250/.00001/.00025 = 10.0×10¹⁰1284 1284-24