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I have a Seurat object

I want to plot a list of genes but I got a warning saying most of genes removed

I have tried with both my genes of interest and all variable genes but the results is very similar

Does this mean these genes are basically are not expressed in my data?

   > t$V1
  [1] "IGH"          "IGHV1-2"      "IGHV1-3"      "IGHV1-12"     "IGHV1-8"     
  [6] "IGHV1-14"     "IGHV1-17"     "IGHV1-18"     "IGHV1-24"     "IGHV1-45"    
 [11] "IGHV1-46"     "IGHV1-58"     "IGHV1-67"     "IGHV1-68"     "IGHV1-69"    
 [16] "IGHV1-69D"    "IGHV1-38-4"   "IGHV1-69-2"   "IGHV2-5"      "IGHV2-10"    
 [21] "IGHV2-26"     "IGHV2-70"     "IGHV2-70D"    "IGHV3-6"      "IGHV3-7"     
 [26] "IGHV3-9"      "IGHV3-11"     "IGHV3-13"     "IGHV3-15"     "IGHV3-16"    
 [31] "IGHV3-19"     "IGHV3-20"     "IGHV3-21"     "IGHV3-22"     "IGHV3-23"    
 [36] "IGHV3-25"     "IGHV3-29"     "IGHV3-30"     "IGHV3-30-2"   "IGHV3-30-3"  
 [41] "IGHV3-30-5"   "IGHV3-32"     "IGHV3-33"     "IGHV3-33-2"   "IGHV3-35"    
 [46] "IGHV3-36"     "IGHV3-37"     "IGHV3-38"     "IGHV3-41"     "IGHV3-42"    
 [51] "IGHV3-43"     "IGHV3-43D"    "IGHV3-47"     "IGHV3-48"     "IGHV3-49"    
 [56] "IGHV3-50"     "IGHV3-52"     "IGHV3-53"     "IGHV3-54"     "IGHV3-57"    
 [61] "IGHV3-60"     "IGHV3-62"     "IGHV3-63"     "IGHV3-64"     "IGHV3-65"    
 [66] "IGHV3-66"     "IGHV3-71"     "IGHV3-72"     "IGHV3-73"     "IGHV3-74"    
 [71] "IGHV3-75"     "IGHV3-76"     "IGHV3-79"     "IGHV3-38-3"   "IGHV3-69-1"  
 [76] "IGHV4-4"      "IGHV4-28"     "IGHV4-30-1"   "IGHV4-30-2"   "IGHV4-30-4"  
 [81] "IGHV4-31"     "IGHV4-34"     "IGHV4-39"     "IGHV4-55"     "IGHV4-59"    
 [86] "IGHV4-61"     "IGHV4-80"     "IGHV4-38-2"   "IGHV5-51"     "IGHV5-78"    
 [91] "IGHV5-10-1"   "IGHV6-1"      "IGHV7-4-1"    "IGHV7-40"     "IGHV7-27"    
 [96] "IGHV7-56"     "IGHV7-34-1"   "IGHV7-81"     "IGHVII-1-1"   "IGHVII-15-1" 
[101] "IGHVII-20-1"  "IGHVII-22-1"  "IGHVII-26-2"  "IGHVII-28-1"  "IGHVII-30-1" 
[106] "IGHVII-30-21" "IGHVII-31-1"  "IGHVII-33-1"  "IGHVII-40-1"  "IGHVII-43-1" 
[111] "IGHVII-44-2"  "IGHVII-46-1"  "IGHVII-49-1"  "IGHVII-51-2"  "IGHVII-53-1" 
[116] "IGHVII-60-1"  "IGHVII-62-1"  "IGHVII-65-1"  "IGHVII-67-1"  "IGHVII-74-1" 
[121] "IGHVII-78-1"  "IGHVIII-2-1"  "IGHVIII-5-1"  "IGHVIII-5-2"  "IGHVIII-11-1"
[126] "IGHVIII-13-1" "IGHVIII-16-1" "IGHVIII-22-2" "IGHVIII-25-1" "IGHVIII-26-1"
[131] "IGHVIII-38-1" "IGHVIII-44"   "IGHVIII-47-1" "IGHV8-51-1"   "IGHVIII-67-2"
[136] "IGHVIII-67-3" "IGHVIII-67-4" "IGHVIII-76-1" "IGHVIII-82"   "IGHVIV-44-1" 
[141] "IGHD1-1"      "IGHD1-7"      "IGHD1-14"     "IGHD1-20"     "IGHD1-26"    
[146] "IGHD2-2"      "IGHD2-8"      "IGHD2-15"     "IGHD2-21"     "IGHD3-3"     
[151] "IGHD3-9"      "IGHD3-10"     "IGHD3-16"     "IGHD3-22"     "IGHD4-4"     
[156] "IGHD4-11"     "IGHD4-17"     "IGHD4-23"     "IGHD5-5"      "IGHD5-12"    
[161] "IGHD5-18"     "IGHD5-24"     "IGHD6-6"      "IGHD6-13"     "IGHD6-19"    
[166] "IGHD6-25"     "IGHD7-27"     "IGHJ1"        "IGHJ1P"       "IGHJ2"       
[171] "IGHJ2P"       "IGHJ3"        "IGHJ3P"       "IGHJ4"        "IGHJ5"       
[176] "IGHJ6"        "IGHA1"        "IGHA2"        "IGHG1"        "IGHG2"       
[181] "IGHG3"        "IGHG4"        "IGHGP"        "IGHD"         "IGHE"        
[186] "IGHEP1"       "IGHM"        
> pbmc=ScaleData(
+     pbmc,
+     features = t$V1)
Centering and scaling data matrix
  |================================================================================| 100%
> DoHeatmap(pbmc,t$V1,group.by = "Condition")
Warning message:
In DoHeatmap(pbmc, t$V1, group.by = "Condition") :
  The following features were omitted as they were not found in the scale.data slot for the SCT assay: IGHEP1, IGHE, IGHJ6, IGHJ5, IGHJ4, IGHJ3P, IGHJ3, IGHJ2P, IGHJ2, IGHJ1P, IGHJ1, IGHD7-27, IGHD6-25, IGHD6-19, IGHD6-13, IGHD6-6, IGHD5-24, IGHD5-18, IGHD5-12, IGHD5-5, IGHD4-23, IGHD4-17, IGHD4-11, IGHD4-4, IGHD3-22, IGHD3-16, IGHD3-10, IGHD3-9, IGHD3-3, IGHD2-21, IGHD2-15, IGHD2-8, IGHD2-2, IGHD1-26, IGHD1-20, IGHD1-14, IGHD1-7, IGHD1-1, IGHVIV-44-1, IGHVIII-82, IGHVIII-76-1, IGHVIII-67-4, IGHVIII-67-3, IGHVIII-67-2, IGHV8-51-1, IGHVIII-47-1, IGHVIII-44, IGHVIII-38-1, IGHVIII-26-1, IGHVIII-25-1, IGHVIII-22-2, IGHVIII-16-1, IGHVIII-13-1, IGHVIII-11-1, IGHVIII-5-2, IGHVIII-5-1, IGHVIII-2-1, IGHVII-78-1, IGHVII-74-1, IGHVII-67-1, IGHVII-65-1, IGHVII-62-1, IGHVII-60-1, IGHVII-53-1, IGHVII-51-2, IGHVII-49-1, IGHVII-46-1, IGHVII-44-2, IGHVII-43-1, IGHVII-40-1, IGHVII-33-1, IGHVII-31-1, IGHVII-30-21, IGHVII-30-1, IGHVII-28-1, IGHVII-26-2, IGHVII-22-1, IGHVII-20-1, IGHVII-15-1, IGHVII-1-1, IGH [... truncated]
> pbmc=ScaleData(
+     pbmc,
+     features =rownames(pbmc))
Centering and scaling data matrix
  |================================================================================| 100%
> rm(pbmc)
> pbmc=readRDS("pbmc.rds")
> pbmc=ScaleData(
+     pbmc,
+     features =rownames(pbmc))
Centering and scaling data matrix
  |================================================================================| 100%
> DoHeatmap(pbmc,t$V1,group.by = "Condition")
Warning message:
In DoHeatmap(pbmc, t$V1, group.by = "Condition") :
  The following features were omitted as they were not found in the scale.data slot for the SCT assay: IGHEP1, IGHE, IGHJ6, IGHJ5, IGHJ4, IGHJ3P, IGHJ3, IGHJ2P, IGHJ2, IGHJ1P, IGHJ1, IGHD7-27, IGHD6-25, IGHD6-19, IGHD6-13, IGHD6-6, IGHD5-24, IGHD5-18, IGHD5-12, IGHD5-5, IGHD4-23, IGHD4-17, IGHD4-11, IGHD4-4, IGHD3-22, IGHD3-16, IGHD3-10, IGHD3-9, IGHD3-3, IGHD2-21, IGHD2-15, IGHD2-8, IGHD2-2, IGHD1-26, IGHD1-20, IGHD1-14, IGHD1-7, IGHD1-1, IGHVIV-44-1, IGHVIII-82, IGHVIII-76-1, IGHVIII-67-4, IGHVIII-67-3, IGHVIII-67-2, IGHV8-51-1, IGHVIII-47-1, IGHVIII-44, IGHVIII-38-1, IGHVIII-26-1, IGHVIII-25-1, IGHVIII-22-2, IGHVIII-16-1, IGHVIII-13-1, IGHVIII-11-1, IGHVIII-5-2, IGHVIII-5-1, IGHVIII-2-1, IGHVII-78-1, IGHVII-74-1, IGHVII-67-1, IGHVII-65-1, IGHVII-62-1, IGHVII-60-1, IGHVII-53-1, IGHVII-51-2, IGHVII-49-1, IGHVII-46-1, IGHVII-44-2, IGHVII-43-1, IGHVII-40-1, IGHVII-33-1, IGHVII-31-1, IGHVII-30-21, IGHVII-30-1, IGHVII-28-1, IGHVII-26-2, IGHVII-22-1, IGHVII-20-1, IGHVII-15-1, IGHVII-1-1, IGH [... truncated]
>
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1 Answer 1

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Specify your genes of interest using the features parameter of the ScaleData() function. If this parameter is not specified, only "variable genes" are scaled and that makes sense, genes that do not demonstrate variability across conditions is not interesting in general. Anyhow, since the DoHeatmap() would be looking at the "scaled data slot", it will not be able to find your genes of interest unless they show variation.

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  • $\begingroup$ Thank you. I use sctranscform, I should scale my data before or after sctransform function? $\endgroup$
    – Mahta Mira
    Aug 5, 2021 at 17:53
  • $\begingroup$ From their vignette: Note that this single command replaces NormalizeData(), ScaleData(), and FindVariableFeatures(). $\endgroup$
    – haci
    Aug 5, 2021 at 17:57
  • $\begingroup$ I checked sctransform with or without scale data and the results was different $\endgroup$
    – Mahta Mira
    Aug 5, 2021 at 18:02
  • 1
    $\begingroup$ 1- Please do not edit your questions extensively, these questions meant to help future users too, now the answer and the question does not match. 2- Please read documentation associated with the functions/packages you use: SCTransform() doc says return.only.var.genes: If set to TRUE the scale.data matrices in output assay are subset to contain only the variable genes; default is TRUE. 3- Please show some effort, you will learn and develop faster if you learn by doing instead of asking questions on each step. $\endgroup$
    – haci
    Aug 5, 2021 at 18:28

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