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XL10 competent cell
Catalog # C503
Introduction
This product is super competent E.coli XL10 cell for chemical transformation with transformation
efficiency greater than 108 cfu/ μg. It can be widely used in plasmid transformation, gene cloning
and blue-white screening.
Package information
Component
C503-01
C503-02
C503-03
XL10 competent cell*
5× 100 μl
10× 100 μl
20× 100 μl
Genetype: Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lacTet r Hte
[F′proAB lacIqZΔM15 Tn10(Tet r) Amy Cam r]
Storage
Stored at -80℃. Do not store the product at -20℃ or in liquid nitrogen.
Advantages
- XL10 strain of E.coli is a kind of new strain for gene cloning with tetracycline (Tet r) and
chloramphenicol (Cam r) resistance.
- Applicable to high efficient transformation of PCR product, cDNA and other unmethylated
DNA.
- EndA1 and RecA1 mutation guarantee the stability of exogenous DNA in the host bacteria,
which is conductive to DNA extraction to get high-quality plasmid.
- Due to the presence of the Hte genotype, the transformation efficiency of large fragments of
DNA is greater, and the growth of bacteria is greatly improved.
- Applicable to blue-white screening of recombinant plasmids.
- Transformation efficiency ≥ 108 cfu/ μg Puc19 DNA.
Quality Control
No exogenous residual plasmid DNA
Transformation efficiency ≥ 108 cfu/ μg Puc19 DNA
Protocols
1. Remove the competent cells from refrigerator of -80℃, and place the competent cells in an ice
water bath to melt.
2. The DNA is added into 100μl competent cells, mix gently. Keep in the ice bath for 30 min.
3. Incubate cells for 90 sec. at 42℃. And return to the ice bath for 2~3 min.
4. Add SOC or LB medium up to a final volume of 1 ml. keep in water bath for 10 min at 37℃.
5. Incubate by shaking (150 rpm) for 45 min at 37℃ to recovery and gain resistance.
6. Centrifuged at 2500g for 5 min, and remove 900 μl supernatant, mix gently. And plate on
selective media. Incubate overnight at 37℃.
Notes
1.
2.
3.
4.
The competent cells should be used immediately after thawed in an ice water bath.
The volume of DNA sample should not exceed 1/10 of the volume of competent cells.
After added the DNA sample, the competent cell should not be pipetting, and just mix gently.
To ensure the high transformation efficiency, the whole operation should be gentle in cool
environment.
Appendix I: the working concentration of common antibiotic.
Antibiotic
Working concentration
Ampicillin
Carbenicillin
Chloramphenicol
Kanamycin
Streptomycin
Tetracycline
100 μg/ml
100 μg/ml
33 μg/ml
30 μg/ml
25 μg/ml
15 μg/ml
Appendix II: the transformation inhibitors in DNA sample and removal methods
Inhibitor of transformation
Removal methods
Detergents
Phenol
Ethanol or isopropanol
PEG
Ethanol precipitation
Chloroform extraction and ethanol precipitation
Dry thoroughly before DNA is dissolved
Column purification or Chloroform extraction and ethanol
precipitation
Column purification or Chloroform extraction and ethanol
precipitation
DNA binding proteins