1. in the prepared library, only 5' P5 -> P7 3' strands bind to the flow cell and 5' P7 -> P5 3' strands do not. correct?
2. in a dual index paired sequencing, the order of sequencing is Read 1, i7 index, i5 index, and then Read 2. it needs three or four sequencing primers. One is for Read 1. One is for i7. Does sequencing i5 need a sequencing primer since the synthesis is the extension from P5 adaptor binding oligo? The oligo can serve as the sequencing primer. The sequencing primer for Read 2 is the reverse complement of the i7 sequencing primer. Correct?
3. An Illumina sequencing kit contains many sequencing primers to fit different library preparation kits. Correct?
4. The reason to do adaptor trimming is that sequencing goes into the sequencing primer region, index region, and adaptor region because the insert is short. For the large fragment, sequencing stops before it goes into the regions. Correct? The adaptor trimming is to trim the sequencing primer region and it's 3' downstream. Correct?
2. in a dual index paired sequencing, the order of sequencing is Read 1, i7 index, i5 index, and then Read 2. it needs three or four sequencing primers. One is for Read 1. One is for i7. Does sequencing i5 need a sequencing primer since the synthesis is the extension from P5 adaptor binding oligo? The oligo can serve as the sequencing primer. The sequencing primer for Read 2 is the reverse complement of the i7 sequencing primer. Correct?
3. An Illumina sequencing kit contains many sequencing primers to fit different library preparation kits. Correct?
4. The reason to do adaptor trimming is that sequencing goes into the sequencing primer region, index region, and adaptor region because the insert is short. For the large fragment, sequencing stops before it goes into the regions. Correct? The adaptor trimming is to trim the sequencing primer region and it's 3' downstream. Correct?
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