TY - JOUR
T1 - A method for isolating and analyzing human mRNA from newborn stool
AU - Bennett, William E.
AU - González-Rivera, Rosbel
AU - Shaikh, Nurmohammad
AU - Magrini, Vincent
AU - Boykin, Michelle
AU - Warner, Barbara B.
AU - Hamvas, Aaron
AU - Tarr, Phillip I.
N1 - Funding Information:
This research has been supported by NIH grants NIDDK T32 DK077653-17 , UL1 RR024992 , UH2 AI083265 , the Institute of Clinical and Translational Sciences (ICTS) at Washington University, and the Doris Duke Clinical Research Fellowship Program. We thank David Hunstad, M.D. for providing human leukocyte RNA samples for use as controls, Rebecca Rashid, Ph.D. for technical advice, and Elizabeth Wolf for administrative assistance.
PY - 2009/9/30
Y1 - 2009/9/30
N2 - Efforts to characterize the human transcriptome have largely been limited to blood, urine, and tissue analyses (i.e., normally sterile materials). We report here an extraction protocol using commercially available reagents to obtain high-yield, reverse-transcribable RNA from human stool. Quantitative reverse transcriptase polymerase chain reactions demonstrated minimal intra-specimen but considerable intra-subject variability over time of transcripts for interleukin-6 (IL-6), IL-8, epidermal growth factor (EGF), calprotectin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This technique now expands opportunities to use the human fecal transcriptome to characterize gastrointestinal pathophysiology.
AB - Efforts to characterize the human transcriptome have largely been limited to blood, urine, and tissue analyses (i.e., normally sterile materials). We report here an extraction protocol using commercially available reagents to obtain high-yield, reverse-transcribable RNA from human stool. Quantitative reverse transcriptase polymerase chain reactions demonstrated minimal intra-specimen but considerable intra-subject variability over time of transcripts for interleukin-6 (IL-6), IL-8, epidermal growth factor (EGF), calprotectin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This technique now expands opportunities to use the human fecal transcriptome to characterize gastrointestinal pathophysiology.
KW - Newborn
KW - Nucleic acid purification
KW - RNA
KW - Stool
KW - Transcriptome
UR - http://www.scopus.com/inward/record.url?scp=70249101695&partnerID=8YFLogxK
U2 - 10.1016/j.jim.2009.07.013
DO - 10.1016/j.jim.2009.07.013
M3 - Article
C2 - 19660464
AN - SCOPUS:70249101695
SN - 0022-1759
VL - 349
SP - 56
EP - 60
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -