From 16b373d2181fc233478cfaa2531b37a36a7d5e25 Mon Sep 17 00:00:00 2001
From: Viktoria Petrova <vipet103@hhu.de>
Date: Wed, 12 Mar 2025 15:20:14 +0100
Subject: [PATCH] add protocol for threedrnaseq

---
 .../protocols/TranscriptQuantification.md     |   2 +-
 assays/ThreeDRNAseq/isa.assay.xlsx            | Bin 18757 -> 16415 bytes
 .../NormalizationWithThreeDRNAseq.md          |   3 +++
 3 files changed, 4 insertions(+), 1 deletion(-)
 create mode 100644 assays/ThreeDRNAseq/protocols/NormalizationWithThreeDRNAseq.md

diff --git a/assays/Salmon_quantification/protocols/TranscriptQuantification.md b/assays/Salmon_quantification/protocols/TranscriptQuantification.md
index e1334ae..6525491 100644
--- a/assays/Salmon_quantification/protocols/TranscriptQuantification.md
+++ b/assays/Salmon_quantification/protocols/TranscriptQuantification.md
@@ -1,3 +1,3 @@
 ## Transcript Quantification with Salmon
 
-To quantify transcripts, all cleaned reads were mapped to the BaRT1 reference (Rapazote-Flores et al., 2019) using Salmon (v. 0.14.1) (Patro et al., 2017), and the mapping rates were estimated with an average of ∼93%. We kept transcripts with a minimum of 1 CPM (counts per million) in at least three samples. Normalization of TPM (transcript per million) was conducted using the ‘ThreeDRNAseq’ R package (v2.0.1) (Guo et al., 2021). For TPM values, see Supplementary Dataset 2 and 3. As the samples were collected at different DAEs and RNA extraction was conducted in several batches, we removed the batch effect in the gene count by using the ThreeDRNAseq’ R package (v2.0.1) (Guo et al., 2021).
\ No newline at end of file
+To quantify transcripts, all cleaned reads were mapped to the BaRT1 reference (Rapazote-Flores et al., 2019) using Salmon (v. 0.14.1) (Patro et al., 2017), and the mapping rates were estimated with an average of ∼93%.
\ No newline at end of file
diff --git a/assays/ThreeDRNAseq/isa.assay.xlsx b/assays/ThreeDRNAseq/isa.assay.xlsx
index 00e33319e8a4ae90081cc6cf516b7015bea118e4..beda1e9f31c63a02d7376f26ed2b4945e17e502f 100644
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diff --git a/assays/ThreeDRNAseq/protocols/NormalizationWithThreeDRNAseq.md b/assays/ThreeDRNAseq/protocols/NormalizationWithThreeDRNAseq.md
new file mode 100644
index 0000000..dec8f61
--- /dev/null
+++ b/assays/ThreeDRNAseq/protocols/NormalizationWithThreeDRNAseq.md
@@ -0,0 +1,3 @@
+## Normalization with ThreeDRNAseq R package
+
+We kept transcripts with a minimum of 1 CPM (counts per million) in at least three samples. Normalization of TPM (transcript per million) was conducted using the ‘ThreeDRNAseq’ R package (v2.0.1) (Guo et al., 2021). For TPM values, see Supplementary Dataset 2 and 3. As the samples were collected at different DAEs and RNA extraction was conducted in several batches, we removed the batch effect in the gene count by using the ThreeDRNAseq’ R package (v2.0.1) (Guo et al., 2021).
\ No newline at end of file
-- 
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