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:8/28/2014)... “This kit has an improved extraction efficiency and provides ... ppb,” said Mark Tess, PhD, Mycotoxin Product Manager for Charm ... and USDA-GIPSA inspection agencies that have a need to detect ... in a matter of minutes with nothing but our equipment ... and grain before accepting it, but they no longer have ...
(Date:8/28/2014)... The ability for organizations to attract top ... individuals to perform at the highest of levels, are ... to compete in this post–recession era. The pharmaceutical ... the sector increasingly focuses on the importance of employee-fit ... real results on the organization's development and the innovation ...
(Date:8/28/2014)... MENLO PARK, Calif. , Aug. 28, 2014 ... company focused on developing novel mid- to late-stage ... raised $7.0 million in its first round of ... AshHill Biomedical Investments and Hercules Bioventures with participation ... biotech entrepreneur and pharmaceutical executive, Dr. Larry ...
(Date:8/28/2014)... 2014 Earlier this month, a ... partnership with the U.S. Air Force on the performance ... indicated that this type of fuel made from mustard ... be important to America’s Eco friendly future. In fact, ... mandate to become independent of foreign oil over the ...
Breaking Biology Technology:Klein Hersh International Leads Philadelphia Area Life Science Industry in Company Culture 2Klein Hersh International Leads Philadelphia Area Life Science Industry in Company Culture 3Armetheon Completes $7.0 Million Series A Financing 2Armetheon Completes $7.0 Million Series A Financing 3Can Small Farmers and a New Seed Based Biofuel “Green” the World? Major Universities and Even the U.S. Air Force Seem to Think So 2Can Small Farmers and a New Seed Based Biofuel “Green” the World? Major Universities and Even the U.S. Air Force Seem to Think So 3Can Small Farmers and a New Seed Based Biofuel “Green” the World? Major Universities and Even the U.S. Air Force Seem to Think So 4Can Small Farmers and a New Seed Based Biofuel “Green” the World? Major Universities and Even the U.S. Air Force Seem to Think So 5
... 24, 2011 Amgen (Nasdaq: AMGN ) today ... full year financial results on Monday, Jan. 24, 2011 after ... followed by a conference call with the investment community at ... will be Kevin Sharer, chairman and chief executive officer, and ...
... (Nasdaq: CEPH ) will hold a public memorial service ... and CEO on February 1, 2011, at 6:00 p.m. at The ... Dr. Baldino founded Cephalon in 1987 and led the ... ten biotechnology companies in the world. He was a recognized pioneer ...
... a strategy-driven biotech and pharmaceutical product development company, announced ... the National Institutes of Health (NIH) Therapeutics for Rare ... and development partner. The RRD and ... development projects: Hereditary Inclusion Body Myopathy (HIBM), a rare ...
Cached Biology Technology:Amgen Announces Webcast of 2010 Fourth Quarter and Full Year Financial Results 2Cephalon to Hold Public Memorial Service to Celebrate the Life of Company Founder, Frank Baldino, Jr., Ph.D. 2RRD Selected as Development Partner for NIH Therapeutics for Rare and Neglected Diseases Program 2RRD Selected as Development Partner for NIH Therapeutics for Rare and Neglected Diseases Program 3
(Date:8/29/2014)... been known about what genetic changes transform wild animals ... of whom is a University of Montana assistant professor, ... the development of the brain and the nervous system ... published Aug. 28 in Science and gives ... http://www.sciencemag.org/ ., The domestication of animals and plants, ...
(Date:8/29/2014)... This news release is available in German . ... of chemical signals between organisms is considered the oldest ... social interactions between conspecifics, for example, the sexual attraction ... trigger social responses and to coordinate reproductive behavior in ... at the University of the Algarve in Faro, Portugal, ...
(Date:8/28/2014)... app called TX Invasives is now available from the ... Texas at Austin for identifying harmful non-native plant, insect ... in the United States cause about $137 billion in ... crops, clog waterways, kill native plants and more. , ... for reporting where invasive species occur, we,re more likely ...
Breaking Biology News(10 mins):New research reveals how wild rabbits were genetically transformed into tame rabbits 2New research reveals how wild rabbits were genetically transformed into tame rabbits 3Ready for mating at the right time 2Ready for mating at the right time 3Mobile app makes ID of harmful plants, insects in Texas a snap 2
... below are the selected highlights for the July 2012 issue ... The July issue is available online at www.genetics.org/content/current ... 2012, Copyright 2012. ISSUE HIGHLIGHTS , ... studies through the use of meta-analysis for structured populations , ...
... Researchers at the Translational Genomics Research Institute (TGen) have developed ... less costly, less invasive and more accurate way of diagnosing ... complication of diabetes, high blood pressure and a form of ... progressive deterioration of the kidney,s ability to filter waste from ...
... satellite imagery have been limited by weather. Clouds, in ... However, University of Iowa scientists have created a ... better estimate the concentration of pollutants, such as soot. ... block remote-sensing satellites, ability to detect, and thus calculate, ...
Cached Biology News:Genetics Society of America's Genetics journal highlights for July 2012 2Genetics Society of America's Genetics journal highlights for July 2012 3TGen method isolates biospecimens for treatment of kidney disease 2UI researchers develop technique to help pollution forecasters see past clouds 2
... Mouse anti-Yersinia pestis F1 antigen, Monoclonal ... ELISA: Use at an ... an assay dependent dilution. WB: Use ... molecular weight: 18 kDa. Not tested ...
Anti-Phospho-JKK/SEK1/MKK4 pAb Description: 100 g rabbit polyclonal antibody. Reacts with human/mouse/rat. Tested in WB. Research Focus: signal transduction Storage: -20C Shipping Tem...
... A - Fixation medium Reagent B - ... for fixing cells in suspension with Reagent ... Reagent B. This procedure gives antibodies access ... scatter characteristics of the cells intact. Specific ...
... formulated for covering tissue sections and cell preparations immunohistochemically stained ... , ... Application: Vision Mount ... that require an aqueous mounting medium, or those that do ...
Biology Products: