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
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Pulsed-field gel electrophoresis with clamped uniform electric field
In clamped homogeneous electric field (CHEF) gel electrophoresis, the electric field is generated by multiple electrodes. These electrodes are arranged in a qua
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
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
Alkaline agarose gel electrophoresis
Alkaline agarose gel electrophoresis is performed at high pH, which causes the loss of a proton from thymine and guanine residues, thus preventing the formation
Rapid isolation of mammalian DNA
Mammalian DNA prepared according to this protocol is approximately 20-50 kb and is suitable for use as a template for PCR reactions. the DNA yield varies betwee
Rapid amplification of cDNA 5' ends (5'-RACE)
There is no greater failure in molecular cloning than the isolation of a cDNA clone that lacks the characteristic structure of the sequence at the 5' end of the
Application of mixed oligonucleotide primer-guided cDNA amplification (MOPAC)
The best way to clone a protein for which only part of the sequence is known is to design an oligonucleotide using the known amino acid sequence and use this ol
polymerase chain reaction (PCR)
The source of this experiment is "Guide to Molecular Cloning Experiments, Third Edition", translated by Huang Peitang et al.