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Ligation in Biological Technology

New Report Just Published: World Endoscopy Systems Market Report

... II-31 Pharyngolaryngoscopy II-31 Obstetric/Gynecology II-32 Fallopian & Ovarian Laparoscopy II-32 Endometriosis II-32 Tubal ligation II-32 Vagina Creation II-32 Laparoscopic Myomectomies II-32 Ovarian Cystectomy II-33 Female Infertility II-33 Female ...

New technique can be breakthrough for early cancer diagnosis

... In 2006 Ola Sderberg and his colleagues at the Department of Genetics and Pathology devised a new technique, in situ PLA (in situ proximity ligation assay), that could detect communication between proteins in cells. These researchers have now refined the method and can now see how proteins ...

High Electrotransformation Efficiencies Obtained With DNA From Ligation Mixtures

... occur when electroporating DNA directly from ligation mixtures, due to components in the ... and heat treatment of the ligation reaction. 2-5 In this study I explored ... for the introduction of DNA directly from ligation reactions. Methods ...

Get the 5'-End

... 5'-end of the gene sequence) by ligation or polymerase extension, and amplifying ... adapter only to the decapped mRNA no ligation occurs to dephosphorylated molecules. Ambion's optimization of both the ligation reaction and the adapter sequence ...

Cloning Based on Efficient Three-Fragment Assembly DNA Ligation

... The efficiency of the Rapid DNA ligation Kit from Roche Molecular Biochemicals was compared with standard and optimized conventional ligation procedures. The reactions generated circular ... simultaneously, yielding circular DNA. Such a ligation is known to require high concentrations of the ...

Molecular Biology:Easy Dephosphorylation and Rapid DNA Ligation

... alkaline phosphatase inactivation, and ligation all in the same tube. ... Ligate DNA in Five Minutes with the Rapid DNA ligation Kit Experience DNA ligation that is fast, flexible, and easy to perform ...

Successful PCR amplification and subcloning of a GC-rich DNA fragment

... was added to 50 ng pDrive Cloning Vector in 1x ligation Master Mix, in a total volume of 10 l. Subsequently, a 3 l aliquot of the ligation reaction was used to transform one tube of QIAGEN ... ampicillin at 30 g/ml. A second set of ligation reactions were carried out, and 3 l aliquots ...

Eppendorf Perfectprep Gel Cleanup Kit

... minutes. Ligation: The ligation reactions were set up based on an insert to ... Transformation: 2 l of each ligation reaction were added to 50 l of the TOP ... The insert and vector in this ligation were purified from agarose using ...

Mammalian Expression Vector for Efficient Cloning of PCR Fragments

... -GCCCGGGC-3 . By including Srf I enzyme in the ligation reaction, the PCR-Script cloning method ... nonphosphorylated, unmodified PCR products. The ligation efficiency of blunt-ended DNA fragments is ... either prokaryotic or eukaryotic systems and (3) ligation of insert to the pCMV-Script vector in the ...

pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning

... DNA polymerase was polished prior to blunt-end ligation to the PCR-Script vector. The PCR product was ... the StrataPrep PCR kit method prior to the ligation reaction. The CAM amplicons were ligated to the ... the amplicons were polished prior to ligation to the PCR Script vector. The cloning efficiency ...

Enhanced PCR Cloning System

... which PCR products are incubated for a 1-hour ligation reaction with one of the Srf I-digested ... 3 . By including the Srf I enzyme in the ligation reaction, the PCR-Script cloning method ... of recircularized vector and increased ligation efficiency for blunt-ended DNA fragments by the ...

Pwo DNA Polymerase

... are blunt-ended and can therefore be used directly for blunt-end ligation without any pretreatment of the ends. For further ... blunt-ended products which can be directly used for blunt-end ligation Note: Pwo DNA Polymerase ...

New Competent Cells for Highest Transformation Efficiencies

... that requires a high efficiency of retrieving transformants from ligation reactions, such as constructing plasmid libraries and PCR cloning. ... volume using 30 ng of vector and 10 ng of library cDNA. Of this ligation reaction, 1 l was transformed into both XL10-Gold and DH10B ...

Generate High-Quality, Directional Plasmid cDNA Libraries

... to construct a complex plasmid library, synthesis of cDNA and ligation to the vector must be as efficient as possible. Ultimately, the ... a plasmid cDNA library is determined by transformation of the ligation products into the E. coli host. The optimal E. coli host for ...

Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence

... vector by LIC, a high-efficiency cloning method that does not require ligation or restriction enzyme digestion. LIC creates a seamless junction 4,5,6 ... the pCAL-n-EK vector is by LIC, a method that does not require ligation or restriction enzyme digestion ( figure 2 ). 4,5,6 ...

Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA)

... methods. It does not require template with long, amplifiable regions, as does MSD. The method does not rely on highly efficient digestion and ligation as does LAP amplification. Finally, GenomePlex has improved on PEP and DOP amplification with the design of improved primers, and does not suffer ...

Electroporation of Yeast Artificial Chromosomes

... low when compared to more recently developed transformation methods. An additional concern is that the smaller YACs of any given ligation mixture are cloned preferentially, therefore prior size fractionation with sucrose gradients to enrich for larger molecules, or ...

Examples of Electro-Transfected Eukaryotic and Prokaryotic Cells

... Influence of DNA size on efficiency Influence of DNA conformation on efficiency Large DNA fragments ligation mixtures Library construction M13 DNA Plasmid incompatibility RNA Release of cell components ...

QIAGEN PCR CloningPlus Kit

... pDrive Cloning Vector 50 ng/l ligation Master Mix 2x solution Distilled water - ...

High-efficiency Derivatives of the BL21 Series for Protein Expression

... EndA + cells very difficult. To retrieve and confirm the plasmid before performing protein expression studies, investigators first transform the ligation mix of the protein expression construct into a high-efficiency, EndA cell line, such as XL1-Blue competent cells. Then, a second transformation ...

Mammalian Expression Vectors with Interchangeable Selectable Markers

... elements without subcloning into such vectors. Cre-Mediated Recombination To eliminate conventional restriction digestion and ligation subcloning procedures, site-specific recombination is used to manipulate DNA fragments. Site-specific recombinases, such as the P1-derived Cre ...

Mammalian and Bacterial Expression in One Vector

... for gel purification of the restriction fragments. Epicurian Coli XL1-Blue supercompetent bacteria were transformed with the ligation product, and 18 colonies were screened by PCR for the presence of the reporter gene. One hundred percent of the screened colonies ...

Using XL1-Red Mutator Strain to Generate Esterase Variants

... are described for the directed evolution of enzymes by random mutagenesis using error-prone PCR or DNA-shuffling. 5,6 For both of these methods, ligation of the PCR products is critical. In addition, the mutation bias must be optimized. We chose to use the mutator strain, Epicurian Coli XL1-Red ...

High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli

... of the desired insert into the pCAL-n-FLAG vector is by ligation-independent cloning (LIC), a method that does not require ligation or restriction enzyme digestion ( Figure 2 , Panel A). The pCAL-n-FLAG vector is pretreated to create noncomplementary, 12- and ...

Restriction Endonucleases Undergo Rigorous QC Testing to Ensure Optimal Performance

... 5 or 3 termini of the fragments, and phosphatases that inhibit ligation reactions by removing the 5 phosphate from DNA ends. To ensure that ... to the reactions is detected as trichloroacetic acid-soluble counts. ligation Assay Figure 1 Stratagene ...

Highest Possible Transformation Efficiencies for High-Throughput Applications

... Hte phenotype are the best choice for any cloning experiment that demands a high efficiency of retrieving transformants from ligation reactions, such as constructing plasmid libraries and PCR cloning. The Hte phenotype helps 96Pack Gold cells fulfill the need ...

Generate Adenovirus Vectors in E. coli by Homologous Recombination with the AdEasy Adenoviral Vector System

... for up to 7.5 kb of foreign DNA. 1 Two methods have traditionally been used to generate recombinant adenoviruses. The first involves direct ligation of the gene of interest into the adenoviral genome. The scarcity of unique restriction sites and the prohibitive size of the genome (36 kb) make ...
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