Hello to everybody!
I am new in Illumina sequencing and have a question (apology if the answer is already somewhere here, but I could not find it)
I had a look on Illumina sequencing primers and the primers which used for library preparation (Nextera) (https://support.illumina.com/content...0002694-01.pdf, http://nextgen.mgh.harvard.edu/attachments/Nextera%20Protocol.pdf) and it seems that they have just 3' end complementarity (19 out of 38 nt). In theory 3' end part would be enough for specific Sequencing by Synthesis. Why do you need this non-complementary 5' tail then? Why don't make it shorter? Or am I just looking at some wrong sequences?
Here are the sequencing primers for the library prepared with Nextera Kit:
Nextera Read 1 Primer: 5′-GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAG-3′
Nextera Read 2 Primer: 5′-GCCTTGCCAGCCCGCTCAGAGATGTGTATAAGAGACAG-3′
Final product ends:
Fw: 5’-AATGATACGGCGACCACCGAGATCTACAC[i5]TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG[GSP part]
Rev: 5’- CAAGCAGAAGACGGCATACGAGAT[i7]GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG[GSP part]
Attached there is a nicer picture
Thanks for help!
I am new in Illumina sequencing and have a question (apology if the answer is already somewhere here, but I could not find it)
I had a look on Illumina sequencing primers and the primers which used for library preparation (Nextera) (https://support.illumina.com/content...0002694-01.pdf, http://nextgen.mgh.harvard.edu/attachments/Nextera%20Protocol.pdf) and it seems that they have just 3' end complementarity (19 out of 38 nt). In theory 3' end part would be enough for specific Sequencing by Synthesis. Why do you need this non-complementary 5' tail then? Why don't make it shorter? Or am I just looking at some wrong sequences?
Here are the sequencing primers for the library prepared with Nextera Kit:
Nextera Read 1 Primer: 5′-GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAG-3′
Nextera Read 2 Primer: 5′-GCCTTGCCAGCCCGCTCAGAGATGTGTATAAGAGACAG-3′
Final product ends:
Fw: 5’-AATGATACGGCGACCACCGAGATCTACAC[i5]TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG[GSP part]
Rev: 5’- CAAGCAGAAGACGGCATACGAGAT[i7]GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG[GSP part]
Attached there is a nicer picture
Thanks for help!
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