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A simple method is described for the efficient recovery of intact mRNA from high resolution agarose gels. Fractionation of RNA is accomplished by gel electrophoresis under denaturing conditions using methylmercuric hydroxide. The RNA in the gel is then transferred electrophoretically to a diethylaminoethyl (DEAE)-membrane. After reversing the methylmercuric modification of the RNA, the membrane is sliced into narrow sections and the RNA is eluted at 65 degrees with a high ionic strength buffer containing 6M guanidine hydrochloride. RNA isolated by this procedure is suitable for subsequent enzymatic reactions, including in vitro translation and reverse transcription. The major advantages offered by this procedure are: 1) The membrane-bound RNA is a replica of the high resolution fractionation pattern achieved in the gel. 2) The immobilization and concentration of RNA and the removal of gel matrix contaminants are all accomplished in one step. 3) Small quantities of RNA are efficiently recovered and are suitable for subsequent biochemical manipulations. The method is of general utility for any biological system. We have applied its use to the fractionation, recovery, and analysis of mRNA from Xenopus liver and have identified cDNA clones complementary to albumin mRNA.
Alwine,
Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
1977, Pubmed
Alwine,
Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
1977,
Pubmed
Bailey,
Methylmercury as a reversible denaturing agent for agarose gel electrophoresis.
1976,
Pubmed
Bittner,
Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets.
1980,
Pubmed
Brosius,
Complete nucleotide sequence of a 23S ribosomal RNA gene from Escherichia coli.
1980,
Pubmed
Brosius,
Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli.
1978,
Pubmed
Buell,
Synthesis of full length cDNAs from four partially purified oviduct mRNAs.
1978,
Pubmed
Danner,
Recovery of DNA fragments from gels by transfer to DEAE-paper in an electrophoresis chamber.
1982,
Pubmed
Deeley,
Primary activation of the vitellogenin gene in the rooster.
1977,
Pubmed
Denhardt,
A membrane-filter technique for the detection of complementary DNA.
1966,
Pubmed
Dunn,
A novel method to map transcripts: evidence for homology between an adenovirus mRNA and discrete multiple regions of the viral genome.
1977,
Pubmed
Goldberg,
Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene.
1980,
Pubmed
Hatfield,
Seryl-tRNA in mammalian tissues: chromatographic differences in brain and liver and a specific response to the codon, UGA.
1970,
Pubmed
Jackson,
Studies on histone organization in the nucleosome using formaldehyde as a reversible cross-linking agent.
1978,
Pubmed
Kutateladze,
New procedure of high-voltage electrophoresis in polyacrylamide gel and its application to the sequencing of nucleic acids.
1979,
Pubmed
Laskey,
Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography.
1975,
Pubmed
Lehrach,
RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination.
1977,
Pubmed
Loening,
The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis.
1967,
Pubmed
Maxam,
Sequencing end-labeled DNA with base-specific chemical cleavages.
1980,
Pubmed
McMaster,
Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.
1977,
Pubmed
Pelham,
An efficient mRNA-dependent translation system from reticulocyte lysates.
1976,
Pubmed
Roberts,
Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ.
1973,
Pubmed
Salim,
Nucleotide sequence of Xenopus laevis 18S ribosomal RNA inferred from gene sequence.
1981,
Pubmed
,
Xenbase
Saris,
Blotting of RNA onto ion exchange paper allowing subsequent characterization by in situ translation in addition to blot hybridization.
1982,
Pubmed
Smith,
Recovery of DNA from gels.
1980,
Pubmed
Southern,
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
1975,
Pubmed
Stellwag,
Electrophoretic transfer of DNA, RNA and protein onto diazobenzyloxymethyl (DBM) - paper.
1980,
Pubmed
Wahl,
Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate.
1979,
Pubmed
Westley,
Xenopus laevis serum albumins are encoded in two closely related genes.
1981,
Pubmed
,
Xenbase
Winberg,
Isolation of DNA from agarose gels using DEAE-paper. Application to restriction site mapping of adenovirus type 16 DNA.
1980,
Pubmed
Zain,
Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA.
1979,
Pubmed
