Setup

Pre-Setup

SampleSheet.csv

You will need to ensure that a SampleSheet is present at the top level of the folder holding the bcl files in your project’s Google bucket. A working SampleSheet.csv is shown below. The demultiplexing should work if you follow this format. Note that the only columns that are strictly necessary (to our knowledge) are sample_id, sample_name, and index.

SampleSheet.csv
[Data],,,,,,,
Sample_ID,Sample_Name,Sample_Plate,Sample_Well,I7_Index_ID,index,I5_Index_ID,index2
1,WT1_Th0_1,,A1,N701,TAAGGCGA,S502,ATAGAGAG
2,WT1_Th0_2,,B1,N701,TAAGGCGA,S503,AGAGGATA
3,WT2_Th0_1,,C1,N701,TAAGGCGA,S504,TCTACTCT
4,WT2_Th0_2,,D1,N701,TAAGGCGA,S505,CTCCTTAC
5,WT3_Th0_1,,E1,N701,TAAGGCGA,S506,TATGCAGT
6,WT3_Th0_2,,F1,N701,TAAGGCGA,S507,TACTCCTT
7,KO1_Th0_1,,A2,N702,CGTACTAG,S502,ATAGAGAG
8,KO1_Th0_2,,B2,N702,CGTACTAG,S503,AGAGGATA
9,KO2_Th0_1,,C2,N702,CGTACTAG,S504,TCTACTCT
10,KO2_Th0_2,,D2,N702,CGTACTAG,S505,CTCCTTAC
11,KO3_Th0_1,,E2,N702,CGTACTAG,S506,TATGCAGT
12,KO3_Th0_2,,F2,N702,CGTACTAG,S507,TACTCCTT
13,WT1_Th1_1,,A3,N703,AGGCAGAA,S502,ATAGAGAG
14,WT1_Th1_2,,B3,N703,AGGCAGAA,S503,AGAGGATA
15,WT2_Th1_1,,C3,N703,AGGCAGAA,S504,TCTACTCT
16,WT2_Th1_2,,D3,N703,AGGCAGAA,S505,CTCCTTAC
17,WT3_Th1_1,,E3,N703,AGGCAGAA,S506,TATGCAGT
18,WT3_Th1_2,,F3,N703,AGGCAGAA,S507,TACTCCTT
19,KO1_Th1_1,,A4,N704,TCCTGAGC,S502,ATAGAGAG
20,KO1_Th1_2,,B4,N704,TCCTGAGC,S503,AGAGGATA
21,KO2_Th1_1,,C4,N704,TCCTGAGC,S504,TCTACTCT
22,KO2_Th1_2,,D4,N704,TCCTGAGC,S505,CTCCTTAC
23,KO3_Th1_1,,E4,N704,TCCTGAGC,S506,TATGCAGT
24,KO3_Th1_2,,F4,N704,TCCTGAGC,S507,TACTCCTT
25,WT1_Th17n_1,,A5,N705,GGACTCCT,S502,ATAGAGAG
26,WT1_Th17n_2,,B5,N705,GGACTCCT,S503,AGAGGATA
27,WT2_Th17n_1,,C5,N705,GGACTCCT,S504,TCTACTCT
28,WT2_Th17n_2,,D5,N705,GGACTCCT,S505,CTCCTTAC
29,WT3_Th17n_1,,E5,N705,GGACTCCT,S506,TATGCAGT
30,WT3_Th17n_2,,F5,N705,GGACTCCT,S507,TACTCCTT
31,KO1_Th17n_1,,A6,N706,TAGGCATG,S502,ATAGAGAG
32,KO1_Th17n_2,,B6,N706,TAGGCATG,S503,AGAGGATA
33,KO2_Th17n_1,,C6,N706,TAGGCATG,S504,TCTACTCT
34,KO2_Th17n_2,,D6,N706,TAGGCATG,S505,CTCCTTAC
35,KO3_Th17n_1,,E6,N706,TAGGCATG,S506,TATGCAGT
36,KO3_Th17n_2,,F6,N706,TAGGCATG,S507,TACTCCTT
37,WT1_Th17p_1,,A7,N707,CTCTCTAC,S502,ATAGAGAG
38,WT1_Th17p_2,,B7,N707,CTCTCTAC,S503,AGAGGATA
39,WT2_Th17p_1,,C7,N707,CTCTCTAC,S504,TCTACTCT
40,WT2_Th17p_2,,D7,N707,CTCTCTAC,S505,CTCCTTAC
41,WT3_Th17p_1,,E7,N707,CTCTCTAC,S506,TATGCAGT
42,WT3_Th17p_2,,F7,N707,CTCTCTAC,S507,TACTCCTT
43,KO1_Th17p_1,,A8,N708,CAGAGAGG,S502,ATAGAGAG
44,KO1_Th17p_2,,B8,N708,CAGAGAGG,S503,AGAGGATA
45,KO2_Th17p_1,,C8,N708,CAGAGAGG,S504,TCTACTCT
46,KO2_Th17p_2,,D8,N708,CAGAGAGG,S505,CTCCTTAC
47,KO3_Th17p_1,,E8,N708,CAGAGAGG,S506,TATGCAGT
48,KO3_Th17p_2,,F8,N708,CAGAGAGG,S507,TACTCCTT
49,WT1_Treg_1,,A9,N709,GCTACGCT,S502,ATAGAGAG
50,WT1_Treg_2,,B9,N709,GCTACGCT,S503,AGAGGATA
51,WT2_Treg_1,,C9,N709,GCTACGCT,S504,TCTACTCT
52,WT2_Treg_2,,D9,N709,GCTACGCT,S505,CTCCTTAC
53,WT3_Treg_1,,E9,N709,GCTACGCT,S506,TATGCAGT
54,WT3_Treg_2,,F9,N709,GCTACGCT,S507,TACTCCTT
55,KO1_Treg_1,,A10,N710,CGAGGCTG,S502,ATAGAGAG
56,KO1_Treg_2,,B10,N710,CGAGGCTG,S503,AGAGGATA
57,KO2_Treg_1,,C10,N710,CGAGGCTG,S504,TCTACTCT
58,KO2_Treg_2,,D10,N710,CGAGGCTG,S505,CTCCTTAC
59,KO3_Treg_1,,E10,N710,CGAGGCTG,S506,TATGCAGT
60,KO3_Treg_2,,F10,N710,CGAGGCTG,S507,TACTCCTT

Configuration

experiment_name

Refers to the common Xavier Lab naming convention of <date_initials>. This will help set the Google bucket location of the input and output files for the experiment; genomics_xavier_bucket/<experiment_type>/<experiment_name>. As is customary, the bcl files that serve as the input to the pipeline must be in genomics_xavier_bucket/<experiment_type>/<experiment_name>/bcl and the fastq files will be written to genomics_xavier_bucket/<experiment_type>/<experiment_name>/fastq.

experiment_type

Denotes the type of experiment that was run. It must correspond to one of the directories listed here, as this parameter is used to set the location for output.

read_pairs_file

Location of the read_pairs.tsv file that will be generated as part of the pipeline.