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NTUni_p5 and NTUni_p7 (these are the Nextera transposes sequences, so their matching sequencing primers will be in the Illumina sequencing kits):
TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG
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CAGAGCACCCGAGCCTCTACACATATTCTCTGTC
Now add the metagenoimcs primers (Hyb_F01_i5 with "ATCACG" and Hyb_R21_i7 with "CGAAAC" (rev comp) in this example) which get incorporated AFTER CYCLE 1:
Cycle 1 - only Hyb_R21_i7 hybridizes and extends to form the reverse compliment of the top strand:
TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG <insert> CTGTCTCTTATACACATCTCCGAGCCCACGAGAC
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CAGAGCACCCGAGCCTCTACACATATTCTCTGTC <insert> GACAGAGAATATGTGTAGACTGCGACGGCTGCT
GGCTCGGGTGCTCTG CAAAGC TAGAGCATACGGCAGAAGACGAAC <- reverse primer binds in cycle 1
Cycle 2 - the newly-formed top strand now has the compliment to the forward primer which can now prime:
AATGATACGGCGACCACCGAGATCTACAC ATCACG TCGTCGGCAGCGTC <- forward primer can now bind
TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG <insert> CTGTCTCTTATACACATCTCCGAGCCCACGAGAC
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AGCAGCCGTCGCAGTCTACACATATTCTCTGTC <insert> GACAGAGAATATGTGTAGACTGCGACGGCTGCTG CAAAGC TAGAGCATACGGCAGAAGACGAAC <- made in cycle 1
GGCTCGGGTGCTCTG CAAAGC TAGAGCATACGGCAGAAGACGAAC <- reverse primer
(this is all happening upside-down to the other strand in cycle 1
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