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Molecular Info® Copy Right © 2001
Institute of Molecular Development LLC
Cloning of a Gene

  • Cloning by mRNA Differential Display
  • Cloning by 5'-RACE
  • Cloning by 3'-RACE
  • Cloning by Genome Walking

  • cDNA Library Construction
  • Screening Lamda Phage cDNA Library
  • Screening Lamda Phage cDNA Expression Library
  • PCR Screening of Lamda Phage cDNA Library
  • DNA Excision from Lamda ZAP Vectors

  • Genomic Library Construction
  • Lamda Phage Vector Preparation for Genomic Library Construction
  • Screening Genomic DNA Library

  • Construction of Combinatorial Peptide Library
  • Construction of Phage Display PeptideLibrary
  • Screening of a Combinatorial Peptide Library
  • Screening of a Phage Display Peptide Library

  • YAC Library Construction
  • Screening of Yeast Two-Hybrid Library
  • Yac Library Screening



  • Electrophoresis
  • Agarose Gel Electrophoresis of DNA
  • DNA Fingerprinting by RFLP and RAPD
  • Genomic DNA Fingerprinting by Pulsed Field Gel Electrophoresis
  • Polyacrylamide Gel Electrophoresis of DNA
  • Preparation of Sequencing Gels
  • Silver staining of DNA

  • Formaldehyde Gel Electrophoresis of RNA
  • Deionization of Formaldehyde, Formamide, and Glyoxal by Mixed Bed Resin
  • Deionization of Formaldehyde, Formamide, and Glyoxal by AG 501-X8 Mixed Bed Resin

  • Native Gel Electrophoresis of Protein
  • SDS-PAGE of Protein
  • Acetic acid-Urea PAGE of Protein
  • 2-D Gel Electrophoresis of Protein
  • Starch Gel Electrophoresis of Protein
  • Coomassie Blue Staining of Protein
  • Silver Staining of Protein



  • Isolation and Manipulation of Nucleic Acids

  • Modification Enzymes
  • Restriction Enzymes

  • DNA Purification from Agarose Gels
  • Removal of RNA from DNA Samples

  • Isolation of Chromosomal DNA from Animal Cells
  • Isolation of Mitochondrial DNA from Animal Cells
  • Isolation of Nuclear DNA from Animal Cells

  • Isolation of Chromosomal DNA from E. coli
  • Isolation of Plasmid DNA
  • Preparation of Phage Lamda DNA
  • Isolation of Yeast DNA
  • Isolation of Fungal DNA

  • Genomic DNA Isolation from Plant Tissues
  • Isolation of Chloroplast and Chloroplast DNA
  • Isolation of Mitochondrial DNA from Cell Suspension Cultures
  • Isolation of Mitochondrial DNA from Plant Tissues
  • Isolation of Mitochondrial DNA from Green Fruits
  • Isolation of Mitochondria for in Organello Protein Synthesis
  • Isolation of Nuclear DNA from Plant Tissues
  • Small Scale Preparation of Genomic DNA from Plant Tissues for PCR

  • Isolation of Poly A+-RNA
  • Isolation of total RNA from paraffin-embedded tissues
  • Removal of DNA from RNA sample
  • Removal of RNase contamination and DEPC-treatment
  • Analysis of RNA quality

  • Isolation of Mitochondrial RNA from Animal Cells
  • Isolation of Total RNA from Animal Cells
  • Isolation of Total RNA from Blood
  • Isolation of Total RNA from Paraffin-Embedded Tissues
  • Isolation of RNA for mRNA Differential Display
  • Isolation of RNA using RNAzol

  • Isolation of Total RNA from Bacterial Cells

  • Isolation of Total RNA from Plant Cells
  • Isolation of Total RNA from Plant Cell Suspension
  • Isolation of Polysomal RNA from Plant Tissues
  • Isolation of Mitochondrial RNA from Plant Cells
  • Isolation of Total RNA from Paraffin-embedded Tissues


    Protein Expression, Purification, and Identification

  • Native Gel Electrophoresis of Protein
  • SDS-PAGE of Protein
  • Acetic acid-Urea PAGE of Protein
  • 2-D Gel Electrophoresis of Protein
  • Starch Gel Electrophoresis of Protein
  • Coomassie Blue Staining of Protein
  • Silver Staining of Protein

  • In vitro Translation Using Wheat Germ
  • Isolation of Mitochondria for in Organello Protein Synthesis

  • Processing of Fusion Proteins
  • Purification of Recombinant Proteins from Inclusion Bodies
  • Purification of Recombinant Proteins Tagged with Affinity Handles
  • Purification of Soluble Recombinant Proteins
  • Purification of Surfactant Protein B
  • Fusion Protein Preparation from Lamda gt11 Phages

  • Protein Purification by Gel Filtration Chromatography
  • Protein Purification by Immunoaffinity Chromatography
  • Radiolabeled Protein Purification by Immunoprecipitation with Antibody-Sepharose
  • Unlabeled Protein Purification by Immunoprecipitation with Antibody-Sepharose
  • Protein Purification by Ion-Exchange Chromatography
  • Protein Purification by Ion-Exchange HPLC
  • Protein Purification by RP-HPLC
  • Protein Purification by Size-Exclusion HPLC
  • Desalting and Concentration of Dilute Proteins
  • Degassing Water, Buffer and Solvent
  • UV Cutoffs of Solvents
  • Physical Properties of Solvents
  • Purification of DNA-Binding Proteins Using Biotin-Streptavidin Affinity Systems
  • Purification of DNA-Binding Proteins Using Biotin-Cellulose Microcolumn
  • Ammonium Sulfate Saturation Table

  • Protein Quantitation by Bradford Method
  • The Lowry Method for Protein Quantitation
  • Protein Quantitation by BCA Assay

  • Identification of Unknown Protein by Mass Spectrometer Analysis


    Polymerase Chain Reaction
  • PCR
  • Long Distance PCR
  • PCR Primers
  • UV Irradiation for De-Contamination
  • RT-PCR
  • Quantitative RT-PCR
  • Semi-Quantitative RT-PCR: Competitive RT-PCR
  • Semi-Quantitative RT-PCR: Noncompetitive RT-PCR
  • In situ PCR
  • In situ RT-PCR
  • PCR in situ Hybridization


    Probe Generation and Labeling
  • Hybridoma Production for Monoclonal Antibody Production
  • Immuno Screening of Hybridomas
  • Purification of Monoclonal Antibodies
  • Production of Polyclonal Antibodies

  • Nonradioactive Labeling of 3'-end Oligonucleotides by Terminal Transferase
  • Nonradioactive Labeling of DNA by Nick Translation
  • Nonradioactive Labeling of DNA by PCR
  • Nonradioactive Labeling of DNA by random priming
  • Nonradioactive Tailling of 3'-end Oligonucleotides by Terminal Transferase
  • Estimating the Yields of Dig-labeled Nucleic Acids
  • RNA Labeling by In vitro transcription

  • 3'-end Labeling of DNA with Klenow Fragment
  • 3'-end Labeling of Oligonucleotides with Terminal Transferase
  • 5'-end Labeling of DNA with T4 Polynucleotide Kinase
  • DNA Labeling by Nick Translation
  • DNA Labeling by PCR
  • DNA Labeling by Random Priming
  • Labeled Single stranded cDNA Synthesis from mRNA
  • Removal of Unincorporated dNTPs

  • RNA Labeling by In vitro Transcription


    Promoter and Genome Analysis

  • DNaseI Finger Printing
  • Gel Mobility Shift Assay for DNA Binding Proteins Using Low-Ionic-Strength PAGE
  • Gel Mobility Shift Assay for DNA Binding Proteins Using High-Ionic-Strength PAGE
  • Purification of DNA Binding Proteins Using Biotin-Streptavidin Affinity Systems
  • Purification of DNA-Binding Proteins Using Biotin-Cellulose Microcolumn
  • Genome Walking by Inverse Polymerase Chain Reaction
  • AFLP (Amplified Fragment Length Polymorphisms)
  • RAPD (Random Amplified Polymorphic DNA)
  • Genomic Fingerprinting by Pulsed Field Gel Electrophoresis


    In vitroMutagenesis
  • Site-directed In vitro Mutagenesis with Oligonucleotides
  • Site-specific In vitro Mutagenesis by Chemical Modifications
  • Gene Targeting by Homologous Recombination
  • Deletion Mutagenesis by Overlap Extension and Splicing PCR
  • Short Sequence-Insertion Mutagenesis by PCR
  • Long Sequence-Insertion Mutagenesis using PCR-Generated Sticky-Feet
  • Random Mutagenesis by PCR


    Sequencing

  • Cycle Sequencing of Purified Plasmid DNA
  • Cycle Sequencing of Bacterial Colonies Containing High-Copy-Number Plasmids
  • Cycle Sequencing of Bacterial Colonies Containing Low-Copy-Number Plasmids
  • Cycle Sequencing of M13, Cosmids, and Lamda DNA
  • Cycle Sequencing of Bacterial Genomic DNA
  • Cycle Sequencing of PCR Products
  • Cycle Sequencing of Genomic DNA
  • Cycle Sequencing by Highly Degenerate Primers
  • Preparation of Sequencing Gels
  • Chemical Sequencing of DNA
  • Dideoxy Chain-Termination Sequencing of plasmid DNA
  • Dideoxy Chain-Termination Sequencing of PCR products


    Signal Transduction

  • In vivo Protein Phosphorylation in Mammalian Cells
  • In vitro Protein Phosphorylation in Mammalian Cell Extracts
  • In vivo Protein Phosphorylation in Plants
  • In vitro Protein Phosphorylation in Plant Cell Extracts


    Signal Detection

  • DNA-DNA Hybridization
  • Alkali Blotting of DNA for Hybridization
  • Tissue Fixation
  • FAA Fixation of Plant Tissue
  • Glutaraldehyde-Osmium Fixation of Plant Tissue
  • Glutaraldehyde-Paraformaldehyde Fixation of Plant Tissue
  • Paraffin Infiltration and Sectioning of Plant Tissue
  • Staining Paraffin Sections
  • Staining Paraffin Sections with Safranin O
  • Staining Paraffin Sections with Safranin-Fast Green
  • Staining Paraffin Sections with Toluidine Blue
  • In situ Hybridization to Metaphase Chromosomes
  • Detection of DNA in Paraffin Embedded Tissue Sections
  • Identification of Bacterial Cells by In situ Hybridization
  • PCR In situ Hybridization
  • In situ PCR
  • In situ Hybridization on Paraffin Embedded Tissues
  • WISH (Whole mount In Situ Hybridization)
  • In situ RT-PCR
  • Western Transfer Blotting
  • Coomassie Blue Staining of Protein
  • Silver Staining of Protein
  • Northern Blot Analysis
  • Ribonuclease Protection
  • PCR-RPA
  • RT-PCR
  • In situ RT-PCR
  • Nuclear Run-On Analysis
  • Chemiluminescent Detection of Digoxigenin-Labeled Probe


    Transformation

  • Calcium Phosphate-Mediated Transformation
  • High Efficiency Calcium Phosphate-Mediated Transformation
  • Gene Transfer by electroporation
  • Gene Transfer by lipofection
  • Gene Transfer by SV40 vectors
  • Transformation by protoplast fusion
  • Gus Reporter Gene Expression in animal tissue

  • One Step Transformation of E. coli
  • Transformation of E. coli
  • Preparation of Competent E.coli cells
  • Transformation of Frozen competent cells
  • Triparental conjugation
  • Insert Screening Cloned in E. coli by PCR

  • Arabidopsis Transformation by Vacuum Infiltration
  • Leaf Disc Transformation
  • Alfalfa Leaf Disc Transformation
  • Tobacco Leaf Disc Transformation

  • Protoplast Preparation
  • Preparation and Culture of Protoplasts from a Sterile Shoot Culture, and Regeneration of Plants
  • Preparation and Culture of Leaf Mesophyll Protoplasts from Greenhouse Grown Plants
  • Preparation and Culture of Protoplasts from Roots of Phaseolus aureus (Mung Bean)
  • Isolation and Culture of Protoplasts from a Cell Suspension Culture of Daucus carota (Carrot)
  • Preparation and Culture of Protoplasts from a Nonmorphogenic Suspension Culture of a Graminaceous Species Lolium multiflorum
  • Protoplast Transformation
  • Transformation of Protoplast-Derived Cells with Agrobacterium
  • Transformation of Protoplast by Chemically Stimulated Uptake of Isolated DNA
  • Transformation of Protoplasts by Fusion of Bacterial Spheroplasts
  • Transformation of Protoplasts by Liposome-Encapsulated Delivery of DNA

  • Gene Targeting in Plants
  • Gus Reporter Gene expression in plants
  • Localization of GUS Reporter Gene Expression in Plants
  • Protein Targeting

  • Yeast Spheroplast Transformation
  • Yeast Transformation by Lithium Chloride


    Biochemical Methods

  • Enzyme assay of UDP-glucuronosyltransferase
  • Preparation of Glutamate from Cell Culture
  • Preparation of Lactate from Cell Culture
  • Preparation of Fatty Acids from Cell Culture


    Cell Biology

  • Isolation of Nuclei from Mammalian Cells
  • Isolation of Microsomes from Mammalian Cells
  • Isolation of Mitochondria from Mammalian Cells
  • Isolation of Nuclei from Plant Tissue
  • Isolation of Microsomes from Plant Tissue
  • Isolation of Microsomes from Fungi
  • Isolation of Chloroplasts
  • Isolation of Mitochondria from Plant Tissue
  • Isolation of Cell Walls from Plant Tissue
  • Flow Cytometric Analysis of Isolated Nuclei from Plant Tissue


    Miscellaneous Information/ Buffers/ Culture Media

  • Biotech Company Addresses
  • Environmentally Friendly Cocktails
  • Radioactive Waste Control
  • Disposal of Ethidium Bromide

  • TE Buffer
  • STE Buffer
  • STET Buffer
  • TNT Buffer
  • Electrophoresis Buffers
  • Enzyme Buffers
  • Gel Loading Buffers

  • LB Medium
  • NZCYM Medium
  • NZYM Medium
  • NZM Medium
  • Terrific Broth
  • SOB Medium
  • SOC Medium
  • 2 X YT Medium
  • M9 Minimal Medium
  • NZY Medium

  • 20% Maltose and 1 M Maltose
  • SM
  • TM
  • Lamda Diluent

  • Physical Properties of Solvents
  • UV Cutoffs of Solvents


    Stock Solutions

  • 30% Acrylamide:Bis Solution
  • 40% Acrylamide:Bis Solution
  • 10 M Ammonium Acetate
  • 10% Ammonium Persulfate
  • Antibiotic Solutions
  • 0.1 M ATP
  • BCIP
  • 2 X BES-Buffered Saline
  • 1 M CaCl2
  • 2.5 M CaCl2
  • 100 mM dNTPs
  • 1 M Dithiothreitol (DTT)
  • 0.5 M EDTA (pH 8.0)
  • Ethidium Bromide (10 mg/ml)
  • 2 X HEPES-Buffered Saline
  • IPTG
  • 1 M Magnesium Acetate
  • 1 M MgCl2
  • 2-Mereaptoethanol (BME)
  • NBT
  • Phenol Preparation
  • Phenol: Chloroform
  • 10 mM Phenylmethylsulfonyl Fluoride (PMSF)
  • Phosphate-Buffered Saline (PBS)
  • 1 M Potassium Acetate (pH 7.5)
  • Potassium Acetate (for Alkaline Lysis)
  • 3 M Sodium Acetate (pH 5.2)
  • 3 M Sodium Acetate (pH 7.0)
  • 5 M NaCl
  • 10% Sodium Dodecyl Sulfate (SDS)
  • 20 X SSC
  • 20 X SSPE
  • Trichloroacetic Acid (TCA)-100% Solution
  • 1 M Tris-HCl
  • Tris-Buffered Saline (TBS) (25 mM Tris)
  • X-Gal
  • Chloroform: IAA
  • 50% Triton X-100
  • 1 M MgSO4
  • 10 ug/ul Glycogen
  • 1 M ZnCl2
  • 1 M Spermidine
  • 0.1 M HEPES/ 1 M HEPES
  • 1 M KCl
  • 5 M NaOH
  • 5% Xylene Cyanol
  • 5% BromoPhenol Blue
  • 10 M LiCl
  • 100 mM NTPs
  • 1 M Sodium Citrate
  • 10% Sarkosyl
  • 50% Nonidet P40
  • 0.5 M EGTA
  • 1 M NaH2PO4
  • 50% Glycerol
  • 100% Denhardt's Solution
  • Salmon Sperm DNA (10 mg/ml)
  • 1 M PIPES (pH 6.8)
  • Proteinase K (20 mg/ml)
  • 1 M Sodium-Phosphate Buffer
  • Pronase (20 mg/ml)
  • RNase A (10 mg/ml)
  • DNase (10 mg/ml)
  • 1 M Tricine-NaOH (pH 8.0-7.0)
  • 1 M Phosphoric Acid
  • 1 M Tris-Acetate (pH 8.0-7.0)
  • 1 M MnCl2
  • 0.5 M Tris-Phosphate (pH 6-8)
  • 1 M Glycine-NaOH Buffer (pH 10)
  • Concentrations of Common Commercial Acids and Bases
  • 1 M (NH4)2SO4
  • 100 mM rATP
  • 1% Gelatin
  • 10% Tween-20
  • 1% Acetylated BSA (10 mg/ml)
  • 1% Toluidine Blue
  • 1% Pepsin
  • 10% Blocking Reagent
  • 3% NiCl2
  • Torula Yeast RNA (10 mg/ml)
  • 1% Coomassie Brilliant Blue R