Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells

Research output: Contribution to journalJournal articlepeer-review

Standard

Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells. / Petersen, N E; Larsen, L K; Nissen, H; Jensen, L G; Jensen, A; Hyltoft Petersen, P; Hørder; Gregersen, N; Kristiansen, K.

In: Clinical Chemistry, Vol. 41, No. 11, 1995, p. 1605-13.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Petersen, NE, Larsen, LK, Nissen, H, Jensen, LG, Jensen, A, Hyltoft Petersen, P, Hørder, Gregersen, N & Kristiansen, K 1995, 'Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells', Clinical Chemistry, vol. 41, no. 11, pp. 1605-13. <http://www.clinchem.org/cgi/content/abstract/41/11/1605>

APA

Petersen, N. E., Larsen, L. K., Nissen, H., Jensen, L. G., Jensen, A., Hyltoft Petersen, P., Hørder, Gregersen, N., & Kristiansen, K. (1995). Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells. Clinical Chemistry, 41(11), 1605-13. http://www.clinchem.org/cgi/content/abstract/41/11/1605

Vancouver

Petersen NE, Larsen LK, Nissen H, Jensen LG, Jensen A, Hyltoft Petersen P et al. Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells. Clinical Chemistry. 1995;41(11):1605-13.

Author

Petersen, N E ; Larsen, L K ; Nissen, H ; Jensen, L G ; Jensen, A ; Hyltoft Petersen, P ; Hørder ; Gregersen, N ; Kristiansen, K. / Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells. In: Clinical Chemistry. 1995 ; Vol. 41, No. 11. pp. 1605-13.

Bibtex

@article{5104ca700f1211de8478000ea68e967b,
title = "Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells",
abstract = "We have improved the protocol for RNA quantification by using RNase protection. Instead of precipitation and extraction with phenol and chloroform, we use a faster and more reliable precipitation based on guanidinium thiocyanate (GdSCN). The internal standard is produced by in vitro transcription of a DNA template constructed so as to allow simultaneous detection of the in vitro transcript and the low-density lipoprotein receptor (LDLR) mRNA by use of the same probe and hybridization. Addition of this internal standard at the step for RNA isolation reduced the analytical imprecision from 40.8% to 19.3%. Estimates of the within- and between-subject biological variations of the LDLR mRNA content in peripheral blood mononuclear cells (PBMCs) isolated from healthy volunteers were 21.5% and 13.6%, respectively, and the analytical imprecision was 22.6%. The mean content of LDLR mRNA in PBMCs from healthy individuals was 0.78 copies per cell.",
author = "Petersen, {N E} and Larsen, {L K} and H Nissen and Jensen, {L G} and A Jensen and {Hyltoft Petersen}, P and H{\o}rder and N Gregersen and K Kristiansen",
note = "Keywords: Guanidines; Humans; Leukocytes, Mononuclear; Liver; Nucleic Acid Hybridization; Precipitation; RNA Probes; RNA, Messenger; Receptors, LDL; Reference Values; Reproducibility of Results; Ribonucleases; Sensitivity and Specificity; Thiocyanates",
year = "1995",
language = "English",
volume = "41",
pages = "1605--13",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry, Inc.",
number = "11",

}

RIS

TY - JOUR

T1 - Improved RNase protection assay for quantifying LDL-receptor mRNA; estimation of analytical imprecision and biological variance in peripheral blood mononuclear cells

AU - Petersen, N E

AU - Larsen, L K

AU - Nissen, H

AU - Jensen, L G

AU - Jensen, A

AU - Hyltoft Petersen, P

AU - Hørder, null

AU - Gregersen, N

AU - Kristiansen, K

N1 - Keywords: Guanidines; Humans; Leukocytes, Mononuclear; Liver; Nucleic Acid Hybridization; Precipitation; RNA Probes; RNA, Messenger; Receptors, LDL; Reference Values; Reproducibility of Results; Ribonucleases; Sensitivity and Specificity; Thiocyanates

PY - 1995

Y1 - 1995

N2 - We have improved the protocol for RNA quantification by using RNase protection. Instead of precipitation and extraction with phenol and chloroform, we use a faster and more reliable precipitation based on guanidinium thiocyanate (GdSCN). The internal standard is produced by in vitro transcription of a DNA template constructed so as to allow simultaneous detection of the in vitro transcript and the low-density lipoprotein receptor (LDLR) mRNA by use of the same probe and hybridization. Addition of this internal standard at the step for RNA isolation reduced the analytical imprecision from 40.8% to 19.3%. Estimates of the within- and between-subject biological variations of the LDLR mRNA content in peripheral blood mononuclear cells (PBMCs) isolated from healthy volunteers were 21.5% and 13.6%, respectively, and the analytical imprecision was 22.6%. The mean content of LDLR mRNA in PBMCs from healthy individuals was 0.78 copies per cell.

AB - We have improved the protocol for RNA quantification by using RNase protection. Instead of precipitation and extraction with phenol and chloroform, we use a faster and more reliable precipitation based on guanidinium thiocyanate (GdSCN). The internal standard is produced by in vitro transcription of a DNA template constructed so as to allow simultaneous detection of the in vitro transcript and the low-density lipoprotein receptor (LDLR) mRNA by use of the same probe and hybridization. Addition of this internal standard at the step for RNA isolation reduced the analytical imprecision from 40.8% to 19.3%. Estimates of the within- and between-subject biological variations of the LDLR mRNA content in peripheral blood mononuclear cells (PBMCs) isolated from healthy volunteers were 21.5% and 13.6%, respectively, and the analytical imprecision was 22.6%. The mean content of LDLR mRNA in PBMCs from healthy individuals was 0.78 copies per cell.

M3 - Journal article

C2 - 7586550

VL - 41

SP - 1605

EP - 1613

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 11

ER -

ID: 11232020