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DH5α Competent Cells for Subcloning
Thermo Scientific™

DH5α Competent Cells for Subcloning

Thermo Scientific™ DH5α Competent Cells for Subcloning are chemically competent E. coli cells recommended for routine subcloning into plasmid vectors.
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Catalog NumberQuantity
EC01114 x 500 μL
Catalog number EC0111
Price (USD)
78.75
Each
Quantity:
4 x 500 μL
Request bulk or custom format
Price (USD)
78.75
Each

Thermo Scientific™ DH5α Competent Cells for Subcloning are chemically competent E. coli cells recommended for routine subcloning into plasmid vectors. Subcloning efficiency cells are not suitable for the generation of cDNA libraries. The φ80dlacZΔM15 marker provides α-complementation of the β-galactosidase gene from pUC or similar vectors to allow blue/white colony screening on bacterial agar plates containing Bluo-Gal or X-Gal.

  • Transformation efficiency: >1x106 cfu/μg pUC19 DNA
  • Suitable for routine cloning applications
  • Convenient economical packaging
  • Contain genetic markers that allow for blue/white colony screening

Applications
DH5α Competent Cells for Subcloning are suitable for the standard DNA cloning applications:

  • DNA cloning following PCR or restriction digestion reactions
  • Subcloning (transferring DNA from a parent vector to a destination vector)
  • Plasmid isolation

Genotype
F– φ80lacZΔ M15 Δ (lacZYA-argF) U169 recA1 endA1 hsdR17 (rK– mK+) phoA supE44 λ- thi–1 gyrA96 relA1

For Research Use Only. Not for use in diagnostic procedures.

Specifications
Bacterial or Yeast StrainDH5α
Blue/White ScreeningYes
Cell TypeChemically Competent
Cloning Methylated DNANo
Contains F' EpisomeLacks F' Episome
Efficiency1x10^6 cfu/μg
For Use With (Application)Cloning
High-throughput CompatibilityNot High-throughput Compatible
Product LineThermo Scientific
Product TypeCompetent Cell
Propagating ccdB VectorsNot for ccdB vector propagation
Quantity4 x 500 μL
Shipping ConditionDry Ice
FormatTube
SpeciesE. coli
Unit SizeEach
Contents & Storage
• 4 x DH5α Competent Cells for Subcloning (500 μL)
• pUC19 DNA (50 μL) (100 pg/μL)

Store at -80°C.

Frequently asked questions (FAQs)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

I'm getting overgrowth of colonies. Why?

Ensure that you are using the correct antibiotic at the appropriate concentration. Additionally, make sure the antibiotic is not expired. If colonies exhibit unexpected morphologies, contamination could be a factor. Check your S.O.C. medium and LB growth medium.

I'm only getting white colonies, but none of the clones have an insert. What can I do?

Here are a few suggestions:

- Small fragments/linkers are cloning in to your vector instead of your insert; to correct this, gel-purify the insert before ligation
- Ensure that the correct concentrations of X-gal and/or IPTG (if vector contains the lacIq marker) are used
- If spreading X-gal and/or IPTG on your plate, allow sufficient time for the reagents to diffuse into the plate
- Incubate your plate for a longer period to ensure full color development

I'm getting no colonies at all on my plates. Can you help?

We recommend trying the following:
- Carry out the puc19 transformation control; this gives you information about the performance of the cells.
- Check plates for expiration and correct media used (LB/agar).
- Confirm that the correct antibiotic and concentration was used.

View all DH5α Competent Cells for Subcloning FAQs