Agilent Technologies E5500 Series Podręcznik Użytkownika Pobierz pdf (Strona 63)

Fragment Library Preparation Using the AB Library Builder

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System: 5500 Series SOLiD

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Systems User Guide

APPENDIXD

Library Construction

Oligonucleotide Sequences

PCR Primer and adaptor sequences

Note: The internal adaptor used for DNA fragment libraries is different from the

internal adaptor used for RNA libraries.

Note:The “~” is a phosphorothioate bond, which protects a sequence from nucleases.

Barcoded adaptor sequences

Adaptor and primer sequences Length (nt)

P1-T Adaptor, 50 µM

5′-CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGA~T-3′

5′-TCACCGACTGCCCATAGAGAGGAAAGCGGAGGCGTAGTGG~C~C-3′

Standard Adaptor, 50 µM

5′-CGCCTTGGCCGTACAGCAG-3′

5′-CTGCCCCGGGTTCCTCATTCTCTGTGTAAGAGGCTGCTGTACGGCCAAGGCGT-3′

Library PCR Primer 1, 50 µM

5′-CCACTACGCCTCCGCTTTCCTCTCTATG-3′ 28

Library PCR Primer 2, 50 µM

5′-CTGCCCCGGGTTCCTCATTCT-3′ 21

Barcoded adaptor sequence Length (nt)

Barcode-T-001, 50 µM

5′-CGCCTTGGCCGTACAGCAG-3′

5′-CTGCCCCGGGTTCCTCATTCTCTGTGTAAGAGGCTGCTGTACGGCCAAGGCGT-3′

Barcode-T-002, 50 µM

5′-CGCCTTGGCCGTACAGCAG3′

5′-CTGCCCCGGGTTCCTCATTCTCTAGGGAGTGGTCTGCTGTACGGCCAAGGCGT-3′

Barcode-T-003, 50 µM

5′-CGCCTTGGCCGTACAGCAG-3′

5′-CTGCCCCGGGTTCCTCATTCTCTATAGGTTATACTGCTGTACGGCCAAGGCGT-3′

1 2 ... 58 59 60 61 62 63 64 65 66 67 68 ... 86 87

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Agilent Technologies E5500 Series Podręcznik Użytkownika Strona 63