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[DNA Library] Introduction of plasmid into Agrobacterium 日期：2005-07-18 00:00:00 点击：113 好评：0
Monday, November 10, 2003 Description Introduction of plasmid into Agrobacterium Procedure Grow 10 ml ON culture at 28C in LB (+25mg/ml gentamycin for GV3101, + 34mg/ml rifampicin for LBA4404) 3. Dilute about 1:100 into 10mL of LB (with ap...
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[DNA Library] Agrobacterium plasmid DNA miniprep 日期：2005-07-18 00:00:00 点击：90 好评：0
Monday, November 10, 2003 Description Agrobacterium plasmid DNA miniprep Procedure 1. Inoculate a single colony into 2 ml of LB or YMB containing appropriate antibiotics. *Set up 3 or 4 cultures and combine the cells. 2. Incubate shaking a...
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[DNA Library] Simplified Arabidopsis Transformation Protocol 日期：2005-07-18 00:00:00 点击：83 好评：0
Friday, November 07, 2003 Description Brief version for those who are familiar with the method Procedure 1.Grow healthy Arabidopsis plants until they are flowering. Grow under long days in pots in soil covered with bridal veil, window scre...
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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：186 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：113 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：93 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：189 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...