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[DNA Library] Quickie Transformations 日期：2005-07-18 00:00:00 点击：151 好评：0
Friday, October 24, 2003 Description We all have to shuffle plasmids/constructs in and out of various E. coli strains for amplification and other purposes. This is a simple five (5) minute protocol for the transformation of E. coli cells....
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[DNA Library] Biotinylation of Antibodies in Phage Display 日期：2005-07-18 00:00:00 点击：157 好评：0
Wednesday, October 22, 2003 Description Biotinylation of Antibodies in Phage Display Procedure 1. Pipette 10 to 50 g of IgG antibody (see Hint #2) into a siliconized microcentrifuge tube (see Hint #3). 2. Add 4.4 l of 1 M NaHCO3 to the IgG...
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[DNA Library] Low Background Phage Display Protocol&nb 日期：2005-07-18 00:00:00 点击：118 好评：0
Description Phage display is a convenient method to select polypeptides that bind to a molecule of interest. The following method is designed to reduce or eliminate the isolation of "background" phage (peptide-displaying phage that bind th...
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[DNA Library] Transfection of Cells with Recombinant&n 日期：2005-07-18 00:00:00 点击：196 好评：0
Description This protocol describes the preparation of electroporation-competent bacteria and their transfection with recombinant bacteriophage DNA. Preparation of a high quality phage display library depends upon a high efficiency of gene...
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[DNA Library] Phage Capture Assay: Quantifying Binding of Receptor to Phag 日期：2005-07-18 00:00:00 点击：114 好评：0
Tuesday, October 21, 2003 Description Two main binding assays can be used to confirm the specific binding of a target receptor (ligate) to phage-borne ligands: the phage capture assay and an ELISA-based procedure described in this protocol...
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[DNA Library] Labeling of DNA Probes using Primer 日期：2005-07-18 00:00:00 点击：94 好评：0
Tuesday, October 21, 2003 Description This technique is an alternative to labeling by nick translation. It is less fussy and generally gives considerably hotter probes. It has the additional advantage of working with even quite dirty DNA pr...
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[DNA Library] Baculovirus Transfection Using Calcium C 日期：2005-07-18 00:00:00 点击：192 好评：0
Description Baculovirus Transfection Using Calcium Chloride Procedure 1. Seed 2 million Sf9 cells into a 25 cm2 T-flask and allow cells to attach for at least 1 hr. 2. Mix 1 ìg of the insect baculovirus vector AcMNPV linear viral DNA and...
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[DNA Library] Crosslinking DNA-Binding Protein to DNA& 日期：2005-07-18 00:00:00 点击：206 好评：0
Sunday, October 19, 2003 Description Crosslinking DNA-Binding Protein to DNA Using a Bromodeoxyuridine-Substituted Probe Procedure 1. Combine 10 g M13 ssDNA vector and an equal molar amount of 17 bp M13 universal primer in a microcentrifuge...
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[DNA Library] -Galactosidase Staining of Eukaryotic Cells in vitro 日期：2005-07-18 00:00:00 点击：169 好评：0
Saturday, October 18, 2003 Description Cells that were previously transfected with a lac Z construct can be stained for -Galactosidase activity. Procedure 1. Wash the cells 2 to 3 times in 2 ml of PBS solution (see Hint #2 and Hint #3). 2....
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[DNA Library] High Efficiency Transformation of Yeast Cells 日期：2005-07-18 00:00:00 点击：134 好评：0
Saturday, October 18, 2003 Description This protocol uses a lithium acetate/PEG treatment to transform yeast with DNA. Plasmid DNA or linear DNA may be used Procedure 1. Use cells from an overnight yeast culture to inoculate 300 ml of liqu...