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@@ -1,7 +1,7 @@
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%% This BibTeX bibliography file was created using BibDesk.
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%% http://bibdesk.sourceforge.net/
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-%% Created for Ryan C. Thompson at 2016-06-10 07:30:41 -0700
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+%% Created for Ryan C. Thompson at 2016-09-24 11:12:07 -0700
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%% Saved with string encoding Unicode (UTF-8)
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@@ -19,10 +19,11 @@
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Year = {2016}}
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@article{globin-reduction,
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+ Abstract = {Primate blood contains high concentrations of globin messenger RNA. Globin reduction is a standard technique used to improve the expression results obtained by DNA microarrays on RNA from blood samples. However, with whole transcriptome RNA-sequencing (RNA-seq) quickly replacing microarrays for many applications, the impact of globin reduction for RNA-seq has not been previously studied. Moreover, no off-the-shelf kits are available for globin reduction in nonhuman primates. Here we report a protocol for RNA-seq in primate blood samples that uses complimentary oligonucleotides to block reverse transcription of the alpha and beta globin genes. In test samples from cynomolgus monkeys (Macaca fascicularis), this globin blocking protocol approximately doubles the yield of informative (non-globin) reads by greatly reducing the fraction of globin reads, while also improving the consistency in sequencing depth between samples. The increased yield enables detection of about 2000 more genes, significantly increases the correlation in measured gene expression levels between samples, and increases the sensitivity of differential gene expression tests. These results show that globin blocking significantly improves the cost-effectiveness of mRNA sequencing in primate blood samples by doubling the yield of useful reads, allowing detection of more genes, and improving the precision of gene expression measurements. Based on these results, a globin reducing or blocking protocol is recommended for all RNA-seq studies of primate blood samples.
},
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Author = {\textbf{Ryan C. Thompson} and Terri Gelbart and Steven R. Head and Phillip Ordoukhanian and Courtney Mullen and Dongmei Han and Dora M. Berman and Amelia Bartholomew and Norma S. Kenyon and Daniel R. Salomon},
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Date-Added = {2016-05-03 23:39:31 +0000},
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- Date-Modified = {2016-05-03 23:44:47 +0000},
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- Journal = {American Journal of Primatology (in review)},
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+ Date-Modified = {2016-09-24 18:11:49 +0000},
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+ Journal = {Journal of Biological Methods (in review)},
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Title = {Optimizing yield of deep {RNA} sequencing for gene expression profiling of peripheral blood samples from cynomolgus monkeys ({Macaca} fascicularis)},
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Year = {2016}}
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