Navigation Links
XL10-Gold Cells Supplied in New Single-Reaction Format


solopack Gold cells reduce the time and effort needed for transformationswithout compromise!

Anna Waesche Jonathan Goodwin Amy Duncan Bruce Jerpseth Alan Greener
Stratagene

Stratagene introduces competent cells from the XL10-Gold strain* in an easy-to-use, single-reaction format that yield the supercompetent efficiency of 1 x 10 9 transformants/g of supercoiled DNA or the competent efficiency of 1 x 10 8 transformants/g of supercoiled DNA. The new Epicurian Coli SoloPack Gold supercompetent and competent cells exhibit the Hte phenotype for high transformation efficiency and exhibit robust growth to produce large colonies. The SoloPack format saves time by providing a convenient transformation format.

Transformation of competent E. coli cells is crucial for cloning a gene, expressing a protein, or constructing a plasmid library. In many of these instances, transformation is the rate-limiting step. Working with an E. coli strain that efficiently transforms ligated DNA in a convenient format is, therefore, desirable. Stratagene developed the XL10-Gold strain, containing the Hte phenotype,** to produce the highest transformation efficiencies for ligated DNA. This phenotype is observed in cells that carry an uncharacterized mutation that has been shown to improve the efficiency of transforming ligated plasmids and large DNA molecules relative to strains of equal competency (as measured by supercoiled pUC DNA transformation).1,2

In any transformation reaction, competent cells must be thawed and aliquotted into Falcon 2059 polypropylene reaction tubes. To speed up the transformation process, we designed the SoloPack format so that the entire transformation reaction, including post heat-shock expression, is performed in a single tube. The SoloPack single-tube, single-reaction format is available at the supercompetent efficiency of 1 x 109 transformants/g of supercoiled DNA and the competent efficiency of 1 x 108 transformants/g of supercoiled DNA.

Although this new format is convenient, it is especially important to maintain high transformation efficiency and not compromise the outcome of the cloning experiment. Hence, we chose our most efficient competent cell strain, XL10-Gold, and created SoloPack Gold supercompetent cells and SoloPack Gold competent cells. Use the supercompetent efficiency (1 x 109 transformants/g of supercoiled DNA) for PCR cloning, restriction enzyme cloning, the ligase-independent cloning (LIC) method,3 and expression vector construction. Or, as an economical alternative, use the competent efficiency of 1 x 108 transformants/g of supercoiled DNA when high efficiency is not required. For plasmid library construction and other demanding experiments where optimal transformation efficiency is critical, Stratagene provides the XL10-Gold ultracompetent cells in our standard-tube format with efficiency greater than 5 x 109 transformants/g of supercoiled DNA.

Transformation

We tested the transformation efficiency of 1 x 109 transformants/g of supercoiled DNA for the SoloPack Gold supercompetent cells: The cells were thawed on ice, 1 l of XL10-Gold b-mercaptoethanol mix was added, and the cells were incubated on ice for 10 minutes. Next, 10 pg of pUC DNA was added to the cells and they were incubated on ice for an additional 20 minutes. The cells were then heat pulsed at different temperatures for varying lengths of time to optimize efficiency. After the heat pulse, 0.15 ml of NZY+ outgrowth media was added, and the cells were shaken in the reaction tubes for 60 minutes prior to plating. Our results showed that a heat pulse of 54C for 60 seconds was optimal to achieve an efficiency of 1 x 109 transformants/g of supercoiled DNA in the single-tube format.

The XL10-Gold strain exhibits a faster growth rate and produces larger colonies than other commonly used hosts like DH5a and XL2-Blue. To quantitate this difference, we performed a standard growth curve for these strains harboring the pBluescript plasmid. Results in Figure 1 show that XL10-Gold cells grew demonstrably faster than DH5a and XL2-Blue cells.

Figure 1

Conclusions

Stratagene now provides SoloPack Gold supercompetent cells (1 x 109 transformants/g supercoiled DNA) and competent cells (1 x 108 transformants/g supercoiled DNA) created from our XL10-Gold strain in a handy single-reaction format. The XL10-Gold strain exhibits the Hte phenotype for high transformation efficiency and provides robust growth to produce larger colonies than other commonly used hosts. Combine the XL10-Gold strain with the SoloPack format, and streamline the transformation process while retaining high efficiency.

Genotype

XL10-Gold strain: Tetr D(mcrA)183 D(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac Hte [F proAB lacI qZDM15 Tn10 (Tetr) Amy Camr]

The XL10-Gold strain is tetracycline and chloramphenicol resistant.

REFERENCES
  1. Jerpseth, B., Callahan, M., and Greener, A. (1997) Strategies 10: 37-38.

  2. Jerpseth, M., et al. (1998) Strategies 11: 3-4.

  3. Aslanidis, C. and de Jong, P.J. (1990) Nucleic Acids Res. 18: 6069-6074.

* U.S. Patent Nos. 5,512,468 and 5,707,841 and patents pending.
** Patent pending.


'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
2. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
3. Cell Proliferation Kit Outperforms the Competition in the Range of Cells Typically Counted
4. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
5. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
6. Simple Isolation of RNA from Tissue and Cultured Cells
7. New BL21-CodonPlus Cells Correct Codon Bias in GC-Rich Genomes
8. TKB1 Cells Identify Receptor Tyrosine Kinase Interacting Proteins
9. Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells
10. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
11. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Gold Cells Supplied New Single Reaction Format

(Date:8/27/2015)... ... August 27, 2015 , ... Inc. Magazine released its annual list of ... number 1,361 in growth for the three years through 2014. Being named to the ... held organizations in the country. , “We are thrilled to make the Inc. ...
(Date:8/26/2015)... Aug. 26, 2015 /PRNewswire/ - iCo Therapeutics ("iCo" or ... financial results for the quarter ended June 30, ... Canadian dollars and presented under International Financial Reporting ... with respect to our Oral Amp B program, ... manufacturer," said Andrew Rae , President & ...
(Date:8/26/2015)... , Aug. 26, 2015 Intrexon Corporation (NYSE: ... today the closing of its previously announced public offering ... the underwriters of their option to purchase an additional ... price of $41.00 per share.  The exercise of the ... common stock sold by Intrexon to 5,609,756 shares and ...
(Date:8/26/2015)... (PRWEB) , ... August 26, 2015 , ... PRC ... to develop new business relationships with local sponsors in southern California. The Clinical Trial ... their drug development processes and regulatory pathways., , Stem ...
Breaking Biology Technology:iLab Solutions Named to 2015 Inc. 5000 List 2iLab Solutions Named to 2015 Inc. 5000 List 3iCo Therapeutics Announces Second Quarter 2015 Financial Results 2iCo Therapeutics Announces Second Quarter 2015 Financial Results 3Intrexon Announces Closing of $230 Million Public Offering of Common Stock including Full Exercise of Underwriters' Option to Purchase Additional Shares 2Intrexon Announces Closing of $230 Million Public Offering of Common Stock including Full Exercise of Underwriters' Option to Purchase Additional Shares 3Intrexon Announces Closing of $230 Million Public Offering of Common Stock including Full Exercise of Underwriters' Option to Purchase Additional Shares 4PRC Clinical To Attend Major Industry Conferences And Meet Sponsor Companies In The San Diego Biotech Hub 2PRC Clinical To Attend Major Industry Conferences And Meet Sponsor Companies In The San Diego Biotech Hub 3
... cells is vital for our bodies to function. Part ... called neurotransmitters. The vesicle fuses with the nerve cell ... the next nerve cell. It is crucial that new ... continuously to take place. If parts of this communication ...
... Cleveland Clinic initiates dosing in ASSERT Trial , TSX ... Resverlogix Corp. ("Resverlogix" or the "Company") (TSX:RVX) announced today ... Phase 2 clinical trial lead by Cleveland Clinic. This ... for the treatment of atherosclerosis, in patients with stable ...
... , SHANGHAI, Dec. 22 ... in China, announced today that,the company has received 2009 Deloitte ... year to win this award. The news was,announced at a ... Kong. , The Deloitte Technology Fast ...
Cached Biology Technology:New insight in nerve cell communication 2Resverlogix Commences Phase 2 Atherosclerosis Clinical Trial 2Resverlogix Commences Phase 2 Atherosclerosis Clinical Trial 3Resverlogix Commences Phase 2 Atherosclerosis Clinical Trial 4
(Date:7/31/2015)... Jul. 31, 2015 NXT-ID, Inc. (NASDAQ: NXTD ... focused on the growing mobile commerce market and creator ... filed provisional patent 62/198989 for ELECTRONIC CRYPTO-CURRENCY ... a method to advance crypto-currencies such as Bitcoin into ... common, uniform way to manage all payments.  ...
(Date:7/30/2015)... , July 30, 2015 Cellecta, Inc., ... gene function analysis and biomarker discovery, announced the ... Knockout Library targeting all human protein coding genes. ... "knock out" a gene,s function. Cellecta,s new Human ... throughput screening tool so that researchers can investigate ...
(Date:7/27/2015)... July 27, 2015   Zynx Health ™, ... clinical improvement solutions, today announced that its ... available on Android smartphones and tablets. With this ... organizations can use ZynxCarebook to securely exchange messages ... streamline care transitions to other care settings, and ...
Breaking Biology News(10 mins):NXT-ID Patents Electronic Crypto-Currency Management Technology 2NXT-ID Patents Electronic Crypto-Currency Management Technology 3NXT-ID Patents Electronic Crypto-Currency Management Technology 4CELLECTA, INC. Announces Launch of Human Whole Genome CRISPR Knockout Library 2Zynx Health Adds Android Device Support to ZynxCarebook Solution 2Zynx Health Adds Android Device Support to ZynxCarebook Solution 3
... at the University of Toronto, The Hospital for Sick ... British Columbia have identified a new treatment target for ... estimated 10% of common colds. The virus, called human ... reason for hospitalization of infants and children under two ...
... August 16th issue of Developmental Cell , researchers at ... that new blood vessels form in the right place and ... long time that blood vessels branch to give rise to ... this branching occurs with a reproducible pattern. However, the mechanisms ...
... A study out of York University has some refreshing ... Published today in the journal Applied Physiology, Nutrition and ... otherwise healthy live just as long as their slim counterparts, ... "Our findings challenge the idea that all obese individuals ...
Cached Biology News:Researchers map pathway of infection for a common, potentially life-threatening respiratory virus 2Fat and healthy? York U study finds slim isn't always superior 2
One-step, microplate or cuvet, colorimetric, detection range 0.1 mg/dL to 3 mg/dL. Procedure: 10 min....
Request Info...
Recombinant Mouse Wnt-3a...
... There are at least 20 structurally ... of the TGF-beta superfamily. BMPs were originally ... bone formation. They are also involved in ... organs. BMPs regulate the growth, differentiation, chemotaxis ...
Biology Products: