Question: demultiplexing with same indices across different lanes
0
gravatar for lait
2.7 years ago by
lait150
lait150 wrote:

When demultiplexing, is it a problem if same indices are used across different lanes?

For example:

  • lane 1 has 4 samples with indices A1, A2, A3 and A4

  • lane 2 has 4 samples with indices A1, A2, A3 and A4

As far as I know, demultiplexing is done per lane? so there should be no problem in this?

ADD COMMENTlink modified 2.7 years ago • written 2.7 years ago by lait150
1
gravatar for genomax
2.7 years ago by
genomax91k
United States
genomax91k wrote:

That is correct. There should be no problem. I assume you are referring to the same indexes being used for different samples.

Note: This will not work with NovaSeq (without XP add-on) or NextSeq series of sequencers. Even though there are optically separate lanes on FC for these sequencers they are fluidically fed by the same sample pool across the entire FC.

ADD COMMENTlink modified 2.7 years ago • written 2.7 years ago by genomax91k

yes correct, different samples across different lanes having same index. thanks for your answer!

ADD REPLYlink written 2.7 years ago by lait150

Please see the Note I added.

ADD REPLYlink written 2.7 years ago by genomax91k

we are using hiseq, I assume there should be no problem then

ADD REPLYlink written 2.7 years ago by lait150

Correct.

ADD REPLYlink written 2.7 years ago by genomax91k

Just one follow up question, should I do a separate demultiplexing sheet for each lane in this case? or is it just one demultiplexing sheet (containing all lanes info, sample names, and indices used) fed to bcl2fastqc, and bcl2fastqc should handle the rest?

ADD REPLYlink modified 2.7 years ago • written 2.7 years ago by lait150

One SampleSheet.csv will be fine. Just make sure you list the samples correctly along with the lane numbers.

ADD REPLYlink modified 2.7 years ago • written 2.7 years ago by genomax91k
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