Exon capture and amplification
This protocol describes the method of exon amplification using the mammalian shuttle vector pSPL3 as an example, which is divided into the following five stages
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Recovery of DNA from agarose and polyacrylamide gels (electroelution to dialysis bags)
This technique (McDonell et al., 1977) allows the recovery of double-stranded DNA from agarose or polyacrylamide gel slices over a wide range of molecular masse
Experiments on purification of radiolabeled oligonucleotides by Sep-Pak C18 column chromatography
This scheme describes a method to separate radiolabeled oligonucleotides from undoped radiolabel using the property of antiphase affinity of oligonucleotides fo
Recovery of DNA from agarose gels (DEAE-cellulose membrane electrophoresis)
Recovery of DNA from agarose gels can be applied to (1) mammalian cell transformation and (2) radiolabeling.
Hybridization experiments with oligonucleotide probes in solution
Jabobs et al. (1988) described the method and principles of hybridization in quaternary ammonium-containing buffers. The following is a simple variation of this
Experiments with oligonucleotides synthesized by labeling with Escherichia coli Klenow fragments
Oligonucleotides are usually labeled using a phosphorylation reaction catalyzed by the T4 phage polynucleotide kinase. However, there is sometimes a need to lab
Purification of RNA from tissues and cells by acid phenol-guanidine thiocyanate-chloroform extraction
The cellular RNAase should be inactivated as soon as possible during the first stage of the extraction process. Once the endogenous RNAase is destroyed, the pos
Pulsed-field gel electrophoresis with transverse alternating fields
Gardiner et al. (1986; Gardiner and Patterson 1989) used a gel electrophoresis device with two sets of platinum wire electrodes placed on either side of a verti
Southern blotting (DNA transfer from one agarose gel to two membranes simultaneously)
DNA can be transferred from both sides of the agarose to both membranes simultaneously. This method is useful when the same restriction endonuclease fragment ne