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AATGATACGGCGACCACCGAAATGATACGGCGACCACCGAGATCTHighlight red red P5 PCR primer/flowcell capture site:
Optional. Example: GATCTTBD. This is called "BarcodeRead2 because it is read after template flip, when Read2 is read. The GSAF does not normally sequence this barcode - please request if you need it read.Highlight yellow yellow BarcodeRead2:
Either the small RNA sequencing primer site: CAGGTTCAGAGTTCTACAGTCCGACGATC OR the standard TruSeq Read 1 primer site: ACACTCTTTCCCTACACGACGCTCTTCCGATCT. Which to chose? The TruSeq Read 1 primer site is complementary to the Read 2 primer site, so if you are designing amplicons do NOT use the TruSeq Read 1 primer site, use the small RNA sequencing primer site.Highlight green green Read1 primer site:
- The insert to be sequenced
Then the Index read primer site: AGATCGGAAGAGCACACGTCTGAACTCCAGTCAC (note the initial A is from the dA tailing of the insert and is not included in the index primer or adaptor sequences; note also the reverse-complement of this is the Read 2 sequencing primer, but the Read 2 sequencing primer includes the T corresponding to the dA insert tail so sequencing starts with the insert)Highlight cyan cyan Read2 primer site:
The index sequence (usually 6 bp)Highlight blue blue BarcodeRead1:
ATCTCGTATGCCGTCTTCTGCTTGHighlight purple purple P7 PCR primer/flowcell capture site:
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