[88] The hot DEAE-column method for transfer RNA fractionation

P. L. Bergquist, B. C. Baguley, R. K. Ralph

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The chapter focuses on the fractionation of transfer ribonucleic acid (tRNA) on columns of diethylaminoethyl (DEAE)-cellulose or DEAE-Sephadex at elevated temperatures that provides a simple procedure for large-scale fractionation of total tRNA or for further separation of tRNA fractions enriched for certain amino acid acceptor species by other methods. The hot column method takes advantage of differences in melting temperatures of various tRNA species, which arise from their different base sequences. Hot column method is used in two ways to fractionate tRNA. Fractionation is achieved at a constant temperature of 65° with a nonlinear salt gradient to elute the tRNA. The second method exploits a decreasing temperature gradient to elute the tRNA at a constant salt concentration. The choice of which of the two methods to use for any particular fractionation depends upon the species of tRNA that are under investigation. The importance of complete removal of ribonuclease from the tRNA prior to hot column chromatography must be emphasized. The fractionation achieved on hot DEAE columns presumably depends upon the degree of unfolding of the various tRNA species at the temperatures and salt concentrations employed. Hence, those tRNA's that are most unfolded are able to form the greatest number of bonds with the DEAE.
LanguageEnglish
Pages660-670
Number of pages11
JournalMethods in Enzymology
Volume12
Issue numberPart A
DOIs
Publication statusPublished - 1967
Externally publishedYes

Fingerprint

Distillation columns
Fractionation
Transfer RNA
Temperature
Salts
Column chromatography
Ribonucleases
Cellulose
Thermal gradients
Freezing
Melting point
Chromatography
Amino Acids

Cite this

Bergquist, P. L. ; Baguley, B. C. ; Ralph, R. K. / [88] The hot DEAE-column method for transfer RNA fractionation. In: Methods in Enzymology. 1967 ; Vol. 12, No. Part A. pp. 660-670.
@article{7411260e830f4d7aabe63f4210d8dd5d,
title = "[88] The hot DEAE-column method for transfer RNA fractionation",
abstract = "The chapter focuses on the fractionation of transfer ribonucleic acid (tRNA) on columns of diethylaminoethyl (DEAE)-cellulose or DEAE-Sephadex at elevated temperatures that provides a simple procedure for large-scale fractionation of total tRNA or for further separation of tRNA fractions enriched for certain amino acid acceptor species by other methods. The hot column method takes advantage of differences in melting temperatures of various tRNA species, which arise from their different base sequences. Hot column method is used in two ways to fractionate tRNA. Fractionation is achieved at a constant temperature of 65° with a nonlinear salt gradient to elute the tRNA. The second method exploits a decreasing temperature gradient to elute the tRNA at a constant salt concentration. The choice of which of the two methods to use for any particular fractionation depends upon the species of tRNA that are under investigation. The importance of complete removal of ribonuclease from the tRNA prior to hot column chromatography must be emphasized. The fractionation achieved on hot DEAE columns presumably depends upon the degree of unfolding of the various tRNA species at the temperatures and salt concentrations employed. Hence, those tRNA's that are most unfolded are able to form the greatest number of bonds with the DEAE.",
author = "Bergquist, {P. L.} and Baguley, {B. C.} and Ralph, {R. K.}",
year = "1967",
doi = "10.1016/S0076-6879(67)12100-9",
language = "English",
volume = "12",
pages = "660--670",
journal = "Methods in Enzymology",
issn = "0076-6879",
publisher = "Academic Press Inc.",
number = "Part A",

}

[88] The hot DEAE-column method for transfer RNA fractionation. / Bergquist, P. L.; Baguley, B. C.; Ralph, R. K.

In: Methods in Enzymology, Vol. 12, No. Part A, 1967, p. 660-670.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - [88] The hot DEAE-column method for transfer RNA fractionation

AU - Bergquist, P. L.

AU - Baguley, B. C.

AU - Ralph, R. K.

PY - 1967

Y1 - 1967

N2 - The chapter focuses on the fractionation of transfer ribonucleic acid (tRNA) on columns of diethylaminoethyl (DEAE)-cellulose or DEAE-Sephadex at elevated temperatures that provides a simple procedure for large-scale fractionation of total tRNA or for further separation of tRNA fractions enriched for certain amino acid acceptor species by other methods. The hot column method takes advantage of differences in melting temperatures of various tRNA species, which arise from their different base sequences. Hot column method is used in two ways to fractionate tRNA. Fractionation is achieved at a constant temperature of 65° with a nonlinear salt gradient to elute the tRNA. The second method exploits a decreasing temperature gradient to elute the tRNA at a constant salt concentration. The choice of which of the two methods to use for any particular fractionation depends upon the species of tRNA that are under investigation. The importance of complete removal of ribonuclease from the tRNA prior to hot column chromatography must be emphasized. The fractionation achieved on hot DEAE columns presumably depends upon the degree of unfolding of the various tRNA species at the temperatures and salt concentrations employed. Hence, those tRNA's that are most unfolded are able to form the greatest number of bonds with the DEAE.

AB - The chapter focuses on the fractionation of transfer ribonucleic acid (tRNA) on columns of diethylaminoethyl (DEAE)-cellulose or DEAE-Sephadex at elevated temperatures that provides a simple procedure for large-scale fractionation of total tRNA or for further separation of tRNA fractions enriched for certain amino acid acceptor species by other methods. The hot column method takes advantage of differences in melting temperatures of various tRNA species, which arise from their different base sequences. Hot column method is used in two ways to fractionate tRNA. Fractionation is achieved at a constant temperature of 65° with a nonlinear salt gradient to elute the tRNA. The second method exploits a decreasing temperature gradient to elute the tRNA at a constant salt concentration. The choice of which of the two methods to use for any particular fractionation depends upon the species of tRNA that are under investigation. The importance of complete removal of ribonuclease from the tRNA prior to hot column chromatography must be emphasized. The fractionation achieved on hot DEAE columns presumably depends upon the degree of unfolding of the various tRNA species at the temperatures and salt concentrations employed. Hence, those tRNA's that are most unfolded are able to form the greatest number of bonds with the DEAE.

UR - http://www.scopus.com/inward/record.url?scp=77956999295&partnerID=8YFLogxK

U2 - 10.1016/S0076-6879(67)12100-9

DO - 10.1016/S0076-6879(67)12100-9

M3 - Article

VL - 12

SP - 660

EP - 670

JO - Methods in Enzymology

T2 - Methods in Enzymology

JF - Methods in Enzymology

SN - 0076-6879

IS - Part A

ER -