Navigation Links
Functional Cloning Using,,,ViraPort Retroviral cDNA Expression Libraries

s human tissues or cell lines by two cycles of purification with oligo dT-cellulose. Double-stranded cDNA was synthesized using Stratagenes cDNA synthesis kit, and size-selected cDNA was inserted between the EcoR I and Xho I sites of the plasmid pFB (Figure 1A). The pFB vector has an extended packaging site and uses the viral splice donor and splice acceptor configuration from the high-titer, high-expressing vector pMFG.7 To minimize packaging size constraints, and because the high infection efficiency of the system obviates the need for selection of transduced cells, there are no selectable markers in this vector. To assess the quality of both the phosphatase-treated EcoR I/Xho I-digested vector as well as the quality of the Daudi cDNA, XL10-Gold ultracompetent cells were transformed with ligated plasmid. Miniprep DNA from 100 of the resultant colonies was digested and analyzed. All of the 100 clones examined had EcoR I/Xho I-excisable inserts, with an average size of 1.7 kb and a size range of 0.5 to 5.0 kb (data not shown). Primary clones (1.1 x 106) were amplified to construct the pFB Daudi library. Figure 2 shows the size distribution for 20 representative cDNAs from this library.

Fig.2

Functional Cloning Strategy

In the schematic showing a general strategy for functional cloning using ViraPort libraries (Figure 1B), high-titer viral stocks were transiently produced by transfecting a 293 cell-based retroviral producer line 8,9 with the plasmid cDNA library, then harvesting cell supernatants after 2 days. Alternatively, 293 or 293T cells (ATCC) may be cotransfected with the plas
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11

Related biology technology :

1. Epitope-Tagging Vectors for Functional Analysis in Yeast
2. The DIG System Nonradioactive Automated High-Throughput In Situ Hybridization: a Powerful Tool for Functional Genomics Research
3. Isolation to Functional Analyses
4. Precursor miRNAs for Successful miRNA Functional Studies
5. Using Validated siRNAs in Functional Genomic Assays
6. Whole Gel Eluter Purification of a Functional Multiprotein DNA Replication Complex, Rev A
7. Sciclone ALH 3000 Liquid Handler: Functional Characterization of the High Volume Head - Part 1: Plate-to-Plate Precision Testing
8. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
9. Enhanced PCR Cloning System
10. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
11. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
Post Your Comments:
(Date:9/30/2014)... Sept. 30, 2014 Report Details ... highest potential revenues Do you want to find ... you revenue predictions for those biological drugs from ... you find financial data, R&D trends, opportunities and ... sales forecasts to 2024 at overall world market, ...
(Date:9/30/2014)... Over the last decade, the laboratory filtration market is ... increased acceptance of new filtration technologies in end users ... also catalyzing the growth of laboratory filtration market. , ... at a strong CAGR during the forecast period of ... by 2019. The market is mainly driven by the ...
(Date:9/30/2014)...  RegeneRx Biopharmaceuticals, Inc. (OTC: RGRX) ("the Company" ... a Canadian Patent for Inhibition or Reversal of ... claims using thymosin beta 4, various fragments and ... purposes.  The patent will expire in 2021. ... ) RegeneRx is focused on ...
(Date:9/30/2014)... Switzerland (PRWEB) September 30, 2014 ... software solutions for drug discovery and related ... FUNGITECT consortium has chosen Genedata Selector as ... sequencing (NGS) data. FUNGITECT is a European ... invasive fungal diseases via individualized anti-fungal drug ...
Breaking Biology Technology:Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 2Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 3Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 4Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 5Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 2Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 3Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 4Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 5RegeneRx Receives Canadian Patent for Inhibition or Reversal of Skin Aging 2FUNGITECT Consortium Adopts Genedata Selector as Data Analysis Platform for Anti-fungal R 2FUNGITECT Consortium Adopts Genedata Selector as Data Analysis Platform for Anti-fungal R 3
... on its technology strengths, will its economic weaknesses eventually ... 21st century knowledge economy are impressive when compared with ... intimidating, especially when it comes to training and retaining ... skilled workers - from construction trades to laboratory technicians ...
... The past is a source of knowledge, and the future is ... S. Ambrose (1936-2002) Ernest Mueller , ... collections represent,a rich source of study material, much of it meticulously ... the field. While the majority of,this material was originally collected and ...
... , , ... NBS microbiologists recently assisted researchers at the University of ... strain of Pichia pastoris yeast. Top,yields by the researchers thus far ... in shake flasks. By switching to a BioFlo 3000 ...
Cached Biology Technology:To be a player in the "Knowledge Economy," Wisconsin needs more tech workers 2To be a player in the "Knowledge Economy," Wisconsin needs more tech workers 3To be a player in the "Knowledge Economy," Wisconsin needs more tech workers 4Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA) 2Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA) 3Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA) 4Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA) 5Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA) 6Genomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Tissues through the use of Whole Genome Amplification (WGA) 7140-Fold Increase of Protein Achieved in Pichia Fermentation 2
(Date:9/30/2014)... German . , ... situations such as exposure to predators. Researchers from the ... long-term study on different populations of great tits that ... temperature. High metabolic rates and low temperatures were associated ... were more likely to approach potential predators. , The ...
(Date:9/30/2014)... suite of perilous threats in today,s ocean. From ... change, fragile coral ecosystems are disappearing at unprecedented ... species of corals surrounding the island of Moorea ... their tropical environment: coral guard-crabs. New research from ... Station scientist Seabird McKeon and the museum,s predoctoral ...
(Date:9/30/2014)... KONG , Sept. 30, 2014 Winners of ... Economist Events, Innovation Summit scheduled to take place at the JW ... the summit, the award winners will share their experiences and the ... who have made a proven innovation over the past decade, will ... the night before the summit. It will be the first ...
Breaking Biology News(10 mins):Risky metabolism 2Smithsonian scientists discover coral's best defender against an army of sea stars 2Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 2Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 3Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 4Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 5Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 6Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 7
... new evidence that certain high blood pressure drugs may be ... of vision loss in people with diabetes. The study, the ... to new ways to prevent or treat the sight-threatening disease, ... Proteome Research , a monthly publication. Edward Feener ...
... Los Angeles, CA Current research suggests that nitric oxide ... The related report by Mittal et al, "Inhibition of ... enhancing phagocytosis of Escherichia coli K1 in neonatal ... American Journal of Pathology . Bacterial meningitis, or ...
... non-neurotoxic treatment for head lice has been found to have an ... be safe in humans from six months of age and up. ... Pediatric Dermatology . Benzyl Alcohol Lotion 5% (known as ... been attempted by treating with household items such as mayonnaise, olive ...
Cached Biology News:Treating neonatal meningitis -- is nitric oxide a foe or a friend to bacteria? 2Study shows that suffocating head lice works in new treatment 2
BD Falcon Center-well 60 mm In Vitro Fertilization (IVF) Dish, nonembryotoxic, noncytotoxic, tissue-culture treated polystyrene. (20/sp, 500/ca) Packaging: 20 / sleeve, 500 / case ...
... Collagen I 60 mm Culture Dishes BD ... treated polystyrene with a uniform application of rat ... Collagen I ,Surface : ECM/attachment ... ,Dim nominal : 60 mm x ...
...
The piston kit includes 2 pistons to service one F10 pump of a BioLogic DuoFlow workstation....
Biology Products: