New procedure of high-voltage electrophoresis in polyacrylamide gel and its application to the sequencing of nucleic acids

Axelrod, V.D.; Gorbulev, V.G.; Belzhelarskaya, S.N.; Vartikyan, R.M.

doi:10.1016/0003-2697(79)90121-0

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Nucleic Acids Research1

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1998

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Cited by 6 publications

(2 citation statements)

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“…The following protocol deviates from previous protocols (13)(14)(15)(16)(17)(18)(19)(20)(21). We built a 38 x 46 x 20 cm transfer device from Plexiglas egg-crate louver panels (from AIN Plastics (Mt.…”

Section: Methodsmentioning

confidence: 99%

Genomic sequencing.

Church¹,

Gilbert²

1984

Proc. Natl. Acad. Sci. U.S.A.

7,567

2,703

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Section: Methodsmentioning

confidence: 99%

Genomic sequencing.

Church¹,

Gilbert²

1984

Proc. Natl. Acad. Sci. U.S.A.

7,567

2,703

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“…The DEAE-membrane is relatively strong and does not disintegrate when wet. It is therefore far more useful than ion-exchange cellulose sheets which have also been used to bind nucleic acids transferred from gels, but which are fragile and very difficult to keep intact (4)(5)(6)(7). Because the DEAE-membrane can be sliced into extremely narrow, yet easily manipulated sections, this recovery procedure preserves the high resolution fractionation of the RNA achieved in the gel to a far greater extent than does any of the methods mentioned above.…”

Section: Nucleic Acids Researchmentioning

confidence: 99%

Efficient recovery of functionally intact mRNA from agarose gels via transfer to an ion-exchange membrane

Holland

Wangh

1983

Nucl Acids Res

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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 650 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.

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Denaturating Electrophoresis in DNA Sequencing: A Brief History and Current Protocols

Kraev

1998

Nucleic Acid Electrophoresis

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New procedure of high-voltage electrophoresis in polyacrylamide gel and its application to the sequencing of nucleic acids

Cited by 6 publications

References 11 publications

Genomic sequencing.

Genomic sequencing.

Efficient recovery of functionally intact mRNA from agarose gels via transfer to an ion-exchange membrane

Denaturating Electrophoresis in DNA Sequencing: A Brief History and Current Protocols

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