TCA-DOC Normal, TCA Acetone Precipitation, Ethanol Precipitation, TCA-DOC / Acetone, Acidified Acetone/Methanol, Dr. Mario Lebendiker. University of Jerusalem. - [Read Protein Precipitation Protocols]
Harvesting and Lysis of Bacteria. Mini-prep plasmid purification using Solution I, Solution II and Solution III. The Preuss Lab. Univ. Chicago. - [Read Alkaline Lysis Mini-Prep Protocol]
Primer design, Primer annealing, Primer labelling, Probe purification, In vitro hybridization DNA-protein Binding reaction. Jonathan Flint. - [Read EMSA Protocol using 32-p.]
Protocol for extraction and purification of total RNA using TRIzol OR TRI reagent. Includes: Homogenization for Cell Suspensions; Phase Separation; RNA Precipitation; RNA Wash; Redissolving the RNA; Determination of RNA Concentration and Purity; Preparation of Rnase-free water. - [Read Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol]
Agarose gel purification,
Annealing and extending, oligonucleotides, Ethanol Precipitation, Ligation
Miniprep, Oligonucleotide purification, Recovering DNA bands, Restriction digest
Gene Clean. The Hu Lab. - [Read DNA cloning Procedures]
PCR polymerase costs can be high. If you are willing to work, you can produce bacteria containing the clone. It appears to produce lots of Taq and is quite stable. The proceedure takes 4 days start to (15 000 units of Taq) finish. The Taq also appears ver - [Read Home-made Taq Polymerase Purification]
From plasmid to protein using bacterial expression. Transform appropriate DNA plasmid, Make a starter culture for protein expression, bacteria culture for protein expression. Sosnick Group Chicago. - [Read Protein Expression and Purification Protocol]
This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography. Mike A. Dyer Lab Harvard. - [Read Antibody Purification]
Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Species and Type of Antibody Agarose
Rabbit IgG Protein A or Protein G
Mouse IgG1 Protein G
Mouse IgG2 Protein A or Protein G
Mouse IgG3 Protein G
Sheep IgG Protein G but binds weakly
Rat IgG Protein G but binds weakly
Guinea Pig IgG Protein A
Dog IgG Protein A
Goat IgG Protein G
Pig IgG Protein A
Hamster IgG Protein G.
By Millipore. - [Read Affinity Antibody Purification of Protein A/G Chromatography]
DNA Extraction from Agarose Gels Protocol. The page includes cutting out the DNA band from the gel, and describes three methods including 1) Spin-columns (Nucleic acid purification columns), 2) using Dialysis tubing (semi-permeable membrane, Visking tubing), and the 3) Paper strip method.Matt Lewis, Department of Pathology, University of Liverpool. - [Read DNA Extraction from Agarose Gels Protocol]
Magnetic DNA affinity purification of yeast transcription factor tau--a new purification principle for the ultrarapid isolation of near homogeneous factor. Gabrielsen et al. 1989 - [Read DNA-Binding Protein Purification with Dynabeads.]
Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Buffer Preparation, Preparation of a Protein A Agarose or Protein G Agarose Affinity Column, Pouring the Protein A/G Affinity Column, Preparation of Antiserum or Ascites for Affinity Chromatography, Affinity Chromatography Using Protein A/G Agarose. - [Read Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography]
Lowry – Protein Determination. Quantitation of total protein using the lowry protein assay. Dr. Mario Lebendiker. The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem. Method materials and notes for the Lowry assay. - [Read Lowry – Protein Determination]
DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.
This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.
This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.