Molecular Biology / Transfection and infection / Virus-mediated transduction protocols and methods

Technique / Molecular Biology / Transfection and infection / Virus-mediated transduction

Herpes simplex virus type-1 HSV amplicon vector construction methods and reviews new recommended protocol
Targeting HSV amplicon vectors (method) An original technique article on HSV vector construction by Grandi and Wang etc. Targeting HSV amplicon vectors. Methods. 2 ...

Transfection Protocol for Vector Production in 293T cells top rated protocol
Lentivirus transfection protocol for vector production in 293T cells from Patrick Salmon's Cookbook, department of Microbiology, Geneva School of Medicine, Switzerland. ...

siRNA Adenovirus Protocol new protocol
Clontech protocol (PDF file) on siRNA adenovirus construction. Useful information can be found from CAGT Vector Development Laboratory, Baylor College of Medicine. ...

Six Well Cell Transfection of 293 Cells protocol
Six Well Cell Transfection of 293 Cells Protocol. (Finkbeiner lab, UCSF) This cell transfection protocol uses Lipofectamine plus Reagent for introducing the plasmid DNA into the cell. ...

Cell Infection with Retrovirus protocol
A retrovirus infection protocol from Shay/Wrigth Laboratory at Southwestern Medical Center. (MS Doc file). ...

Protocol for 293T transfection and viral titering new protocol
Protocol for Plasmid transfection and retrovirus construction in 293 cells. The protocol can be downloaded as word file.(Dr.Gordon Fishell, New York University School of Medicine) ...

A Protocol for adeno-associated AAV vector production and purification new protocol
The protocol describe at below is typical of methods that are used topropagate and purify AAV vectors for experiments both in vitro and in vivo. (Hiroaki Mizukami et al., Jichi Medical School, Japan) ...

HEK 293 cells top rated troubleshooting
HEK 293 cells are Human Embryonic Kidney cells (HEK cells). 293 cells are generated by transformation of sheared adenovirus 5 genome into human embryonic kidney cells. It is one of the most commonly used cell lines because of its high transfection efficiency. ...

Troubleshooting and Optimization of Retroviral Transduction Parameters troubleshooting
One of the most frequent laments of using recombinant retroviruses is that the titer does not appear to be sufficient to infect the cells of interest at a high enough frequency to suit your purposes. This is a troubleshooting guide using Phoenix lines. (No ...

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16-May-2008 04:39 pm

Related new papers and reviews

Retroviral transduction of murine hematopoietic stem cells.

Hematopoietic stem cells (HSC) are inherently rare cell types that cannot be obtained in sufficient amounts for classical biochemical characterization. To facilitate functional studies of murine HSC and hematopoietic development, the technique of retroviral-mediated gene transfer provides a useful tool. The generation of high titer retroviral vectors permits transduction of stem cells with a variety of genes and leads to long-term marking in the blood of recipient mice. Optimized promoter/enhancers facilitate high-level transgene expression in mice transplanted with transduced bone marrow (BM) cells. The co-expression of reporter genes along with a gene of interest greatly facilitates tracking donor engraftment of transduced hematopoietic progeny following stem cell transplantation. This methodology can be used to reconstitute defective function in a mutant background or to study protein function during hematopoiesis by overexpression. Despite limitations such as integration site variegation and copy number-dependent effects, this approach is rapid and efficient compared with transgenic mouse technology. In this chapter, we review this broadly applicable technique for achieving high-level murine BM stem cell transduction. We also describe methods for transplantation and subsequent analysis of transplanted mice as a bona fide assay for the stem cell transduction efficiency.