How to trim adapters from miRNA data sequenced on Illumina machine?
Go Back To trim the small RNA adapter from Illumina microRNA (miRNA) reads please create a Trim Adapter List in the following way:
Why should the Trim adapter list be created this way?
Since the read is sequenced from the 5' end through the miRNA sequence and into the adapter sequence, it is the 3' end which should be trimmed away. If your reads are as in the 36 nt long example. The first nt are the miRNA (21 nt in this example) followed by the adapter (24 nt in this example with Small RNA v1.5 3′Adapter), which then extend beyond the read. Hence, you need to select the option "Allow end trimming". The minimum end score should be according to the specificity what you wish to use for when an adapter is recognized. If you for example set it to 6, as done in our tutorial, you will allow for up to three mismatches or two gaps in cases where the miRNA is 21 nt long. If your reads are longer, say, 50 bp, then the adapter sequence will be found in the middle of the read. The first nt are the miRNA, which in this example is 21 nt miRNA, followed by the 21 nt adapter (TruSeq Small RNA in this example) + 8 nt included after the adapter sequence. If this is the case you will need to select the option "Allow internal matches". The minimum internal score should be according to the specificity what you wish to use for when an adapter is recognized. If leaving the default, which is 10, then it will allow up to three mismatches or two gaps for the 21 bp adapter.
Our tutorial using the 36 bp long reads can be found in our webpage as follows: https://digitalinsights.qiagen.com/clc-genomics-workbench-resources/
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