Dear all,
my lab conducted a sequencing task using MinION. It's the first time I have to analyze data from MinION sequencer. I am confused about the organization of the data produced. the folder tree looks like reported in the picture.
I guess the data I have to start from are contained in the barcode01...barcode12 folders (collection of fast5 files). Please, can someone let me understand:
What's the meaning of the barcodes folders?
Where the fast5 files in the barcode folders come from?
Are the fast5 files in every barcode folder independent?
barcodes are used during the library preparation to "label" different samples. Because each sample has been labeled with a unique barcode, all the fast5 files in one barcoded folder comes from the same sample, i.e. they are not independent.
You can talk to your lab members about the barcodes used and the samples behind them.
The point of barcoding / multiplexing is to increase the number of independent samples we can sequence in one run, where if we use 8 barcodes each sample theoretically should get 1/8 of the total reads.
The samples, and therefore the barcodes as folders, are independent. (else there would be no point in barcoding)
The fast5s within each barcode folder come the same sample, and can be basecalled and the resulting fastq reads combined.
Thank you for your answer.
I asked the lab member give me the experiment summary (you can find it attached).
As I understand (for example), the sample 1 (super-pool) is associated to barcode 1 whose folder contains 178 fast5s. So, to start my analysis, I can run the pipeline to every barcode folder, one-by-one. Is it right?
Emilio
Thank you for your answer. I asked the lab member give me the experiment summary (you can find it attached). As I understand (for example), the sample 1 (super-pool) is associated to barcode 1 whose folder contains 178 fast5s. So, to start my analysis, I can run the pipeline to every barcode folder, one-by-one. Is it right?
Emilio