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Canonical ILLUMINA library design as of Nov. 2011 (all 5'-3'), "TruSeq V3":
- P5 PCR primer/flowcell capture site:AATGATACGGCGACCACCGA
- BarcodeRead2:Optional. Example: GATCT. 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.
- Read1 primer site:Either the small RNA sequencing primer site: CAGGTTCAGAGTTCTACAGTCCGACGATC OR the standard TruSeq Read 1 primer site: ACACTCTTTCCCTACACGACGCTCTTCCGATCT
- The insert to be sequenced
- Read2 primer site: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)
- BarcodeRead1:The index sequence (usually 6 bp)
- P7 PCR primer/flowcell capture site:ATCTCGTATGCCGTCTTCTGCTTG
Barcodes (also known as Indexes)
NOTE: Illumina barcodes (indexes) have varied significantly over time NOT ONLY in their sequence but also in WHERE they are placed in the sequencing construct.
The GSAF expects indexes to be in the 3' end of the final sequencing construct, between the Index read sequencing primer site and the P7 PCR primer site. If you are using dual-indexed samples with an additional barcode between the P5 bridge PCR primer site and the Read 1 sequencing primer site, we can easily accommodate that on a run but do not normally do so - you need to tell us.
The GSAF uses the following names for the following barcodes. Note that these sequences are shown 5'-3' when the P5 sequence is on the right. In other words, here is the first barcode shown in the context of the full 3'-end adaptor construct:
GATCGGAAGAGCACACGTCTGAACTCCAGTCACATCACGATCTCGTATGCCGTCTTCTGCTTG
Sequence |
TruSeq name |
NEXTFlex number |
---|---|---|
ATCACG |
TSBC01 |
NFBC07 |
CGATGT |
TSBC02 |
NFBC01 |
TTAGGC |
TSBC03 |
NFBC08 |
TGACCA |
TSBC04 |
NFBC02 |
ACAGTG |
TSBC05 |
NFBC03 |
GCCAAT |
TSBC06 |
NFBC04 |
CAGATC |
TSBC07 |
NFBC05 |
ACTTGA |
TSBC08 |
NFBC09 |
GATCAG |
TSBC09 |
NFBC10 |
TAGCTT |
TSBC10 |
NFBC11 |
GGCTAC |
TSBC11 |
NFBC12 |
CTTGTA |
TSBC12 |
NFBC06 |
AGTCAA |
TSBC13 |
NFBC13 |
AGTTCC |
TSBC14 |
NFBC14 |
ATGTCA |
TSBC15 |
NFBC15 |
CCGTCC |
TSBC16 |
NFBC16 |
GTAGAG |
TSBC17 |
NFBC17 |
GTCCGC |
TSBC18 |
NFBC18 |
GTGAAA |
TSBC19 |
NFBC19 |
GTGGCC |
TSBC20 |
NFBC20 |
GTTTCG |
TSBC21 |
NFBC21 |
CGTACG |
TSBC22 |
NFBC22 |
GAGTGG |
TSBC23 |
NFBC23 |
GGTAGC |
TSBC24 |
NFBC24 |
ACTGAT |
TSBC25 |
NFBC25 |
ATGAGC |
TSBC26 |
NFBC26 |
ATTCCT |
TSBC27 |
NFBC27 |
CAAAAG |
TSBC28 |
NFBC28 |
CAACTA |
TSBC29 |
NFBC29 |
CACCGG |
TSBC30 |
NFBC30 |
CACGAT |
TSBC31 |
NFBC31 |
CACTCA |
TSBC32 |
NFBC32 |
CAGGCG |
TSBC33 |
NFBC33 |
CATGGC |
TSBC34 |
NFBC34 |
CATTTT |
TSBC35 |
NFBC35 |
CCAACA |
TSBC36 |
NFBC36 |
CGGAAT |
TSBC37 |
NFBC37 |
CTAGCT |
TSBC38 |
NFBC38 |
CTATAC |
TSBC39 |
NFBC39 |
CTCAGA |
TSBC40 |
NFBC40 |
GACGAC |
TSBC41 |
N/A |
GCGCTA |
N/A |
NFBC41 |
TAATCG |
TSBC42 |
NFBC42 |
TACAGC |
TSBC43 |
NFBC43 |
TATAAT |
TSBC44 |
NFBC44 |
TCATTC |
TSBC45 |
NFBC45 |
TCCCGA |
TSBC46 |
NFBC46 |
TCGAAG |
TSBC47 |
NFBC47 |
TCGGCA |
TSBC48 |
NFBC48 |
NOTE that TSBC41 is hamming distance 2 away from both TSBC31 and TSBC11; all others are hamming distance >=3.
Some additional 5 bp barcodes can be found here:http://comailab.genomecenter.ucdavis.edu/index.php/Barcodes
An additional set of 6 bp barcodes that have hamming distance of at least 3 from the table above of 49 barcodes is as follows (NOTE: these have NOT been tested on the sequencer):
Sequence |
GSAF name |
|
---|---|---|
AAACAC |
UTBC50 |
|
TGAAGG |
UTBC51 |
|
AACATA |
UTBC52 |
|
CGCGTC |
UTBC53 |
|
GATACA |
UTBC54 |
|
GGTGTG |
UTBC55 |
|
TAAGAA |
UTBC56 |
|
AGCGAG |
UTBC57 |
|
CGGTTA |
UTBC58 |
|
AGCTTT |
UTBC59 |
|
TGGTCT |
UTBC60 |
|
TATCCC |
UTBC61 |
|
TGTCGT |
UTBC62 |
|
CCCCAC |
UTBC63 |
|
ATACGA |
UTBC64 |
|
CCCTTG |
UTBC65 |
|
ACCGGC |
UTBC66 |
|
TTACTG |
UTBC67 |
|
GGAACT |
UTBC68 |
|
GTTATT |
UTBC69 |
|
AAAAGT |
UTBC70 |
|
AAGGGA |
UTBC71 |
|
AAGTAT |
UTBC72 |
|
ACATCT |
UTBC73 |
|
ACGATT |
UTBC74 |
|
ACGCCG |
UTBC75 |
|
ACTCTC |
UTBC76 |
|
AGAATC |
UTBC77 |
|
ATTGGG |
UTBC78 |
|
CCGCGT |
UTBC79 |
|
CGCCCT |
UTBC80 |
|
CTGCAG |
UTBC81 |
|
GAAGTT |
UTBC82 |
|
GCACCC |
UTBC83 |
|
GCAGGA |
UTBC84 |
|
GCCGCG |
UTBC85 |
|
GGCGGT |
UTBC86 |
|
GTATTA |
UTBC87 |
|
TACGTG |
UTBC88 |
|
TCACAT |
UTBC89 |
|
TCTATA |
UTBC90 |
|
TGCAAA |
UTBC91 |
|
TGGCAC |
UTBC92 |
|
TGTTAG |
UTBC93 |
|
TTCTAT |
UTBC94 |
|
Excruciating details - USE WITH CAUTION - RNA PCR primers are NOT current as of Dec. 2011
Oligonucleotide sequences for TruSeqTM RNA and DNA Sample Prep Kits1 TruSeq Universal Adapter 5’ AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT TruSeq Adapters barcode: ATCACG TruSeq Adapter, Index 1 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACATCACGATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 2 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACCGATGTATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 3 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACTTAGGCATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 4 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACTGACCAATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 5 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACACAGTGATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 6 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACGCCAATATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 7 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACCAGATCATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 8 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACACTTGAATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 9 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACGATCAGATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 10 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACTAGCTTATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 11 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACGGCTACATCTCGTATGCCGTCTTCTGCTTG TruSeq Adapter, Index 12 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACCTTGTAATCTCGTATGCCGTCTTCTGCTTG 5' GATCGGAAGAGCACACGTCTGAACTCCAGTCAC Mux index read seq primer AGATCGGAAGAGCACACGTCTG - my 3' adaptor my RT primer: 5' TCAGACGTGTGCTCTTCCGATCT 3' TCTAGCCTTCTCGTGTGCAGACTTGAGGTCAGTG Mux read 2 seq primer (reversed) CTTGAGGTCAGTGGAACATTAGAGCATACGGCAGAAGACGAAC Index 12 PCR primer (reversed) So order RC of TruSeq Adaptor Indexes as PCR primers Oligonucleotide sequences for TruSeq Small RNA Sample Prep Kits ***************************** Example (all Illumina sequences unless noted): RNA PCR Primer (RP1), part # 15005505 5’ AATGATACGGCGACCACCGAGATCTACACGTTCAGAGTTCTACAGTCCGA AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGTCCGA - this is the NEB SR Primer R1 - pads added by SPHS 5' RNA adaptor GUUCAGAGUUCUACAGUCCGACGAUC (NEB kit calls this the SR Adaptor 1) RNA PCR Primer, Index 1 (RPI1) code ATCACG in DNA, RC (CGTGAT) 5' CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA 3' - RNA pcr primer CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC (RC of DNA Index 1 for comparison) 3' RNA adaptor 3' CCTTGGCACCCGAGAATTCCA - 5' ***************************** Differences with NEB Small RNA kit (RP1/SR Primer R1/SR Adaptor 1/5' RNA adaptor are all the same): 5’ GATCGGAAGAGCACACGTCTGAACTCCAGTCACCTTGTAATCTCGTATGCCGTCTTCTGCTTG (TruSeq Adaptor Ind 12) NEB 3' adaptor (3' SR Adaptor 1) 5' ATCGTATGCCGTCTTCTGCTTG 3' 3' AGCATACGGCAGAAGACGAAC 5' (reverse of NEB SR Primer F1) 3' CTAGCCTTCTCGTGTGCAGACTTGAGGTCAGTGCCGATGTAGAGCATACGGCAGAAGACGAAC 5' (rev. of Ind. 12 adaptor) CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGTTC (TruSeq Adaptor Ind 12 PCR primer) 5' CAAGCAGAAGACGGCATACGA 3' (NEB SR Primer F1) 5' CAAGCAGAAGACGGCATACGAGCTCTTCCGATCT (Illumina genomic PCR primer) 3' CAAGCAGAAGACGGCATACGAT 5' - NEB 3' SR Adaptor 1 (RC) 5' CAAGCAGAAGACGGCATACGA 3' - NEB RT Primer 1 For reference: >TruSeqAdapterIndex11_RC CAAGCAGAAGACGGCATACGAGATGTAGCCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC >TruSeqAdapterIndex12_RC CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC ***************************** RNA 5’ Adapter (RA5), part # 15013205 5’ GUUCAGAGUUCUACAGUCCGACGAUC RNA 3’ Adapter (RA3), part # 15013207 5’ TGGAATTCTCGGGTGCCAAGG 3' ACCTTAAGAGCCCACGGTTCCG 5' (RT primer, reversed) RNA RT Primer (RTP), part # 15013981 5’ GCCTTGGCACCCGAGAATTCCA Multiplexing Index Read Sequencing Primer 5' GATCGGAAGAGCACACGTCTGAACTCCAGTCAC Multiplexing Read 2 Sequencing Primer 5' GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT RNA PCR Primer (RP1), part # 15005505 5’ AATGATACGGCGACCACCGAGATCTACACGTTCAGAGTTCTACAGTCCGA RNA PCR Primer, Index 1 (RPI1) CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 2 (RPI2) CAAGCAGAAGACGGCATACGAGATACATCGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 3 (RPI3) CAAGCAGAAGACGGCATACGAGATGCCTAAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 4 (RPI4) CAAGCAGAAGACGGCATACGAGATTGGTCAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 5 (RPI5) CAAGCAGAAGACGGCATACGAGATCACTGTGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 6 (RPI6) CAAGCAGAAGACGGCATACGAGATATTGGCGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 7 (RPI7) CAAGCAGAAGACGGCATACGAGATGATCTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 8 (RPI8) CAAGCAGAAGACGGCATACGAGATTCAAGTGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 9 (RPI9) CAAGCAGAAGACGGCATACGAGATCTGATCGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 10 (RPI10) CAAGCAGAAGACGGCATACGAGATAAGCTAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 11 (RPI11) CAAGCAGAAGACGGCATACGAGATGTAGCCGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 12 (RPI12) CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 13 (RPI13) CAAGCAGAAGACGGCATACGAGATTTGACTGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 14 (RPI14) CAAGCAGAAGACGGCATACGAGATGGAACTGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 15 (RPI15) CAAGCAGAAGACGGCATACGAGATTGACATGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 16 (RPI16) CAAGCAGAAGACGGCATACGAGATGGACGGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 17 (RPI17) CAAGCAGAAGACGGCATACGAGATCTCTACGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 18 (RPI18) CAAGCAGAAGACGGCATACGAGATGCGGACGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 19 (RPI19) CAAGCAGAAGACGGCATACGAGATTTTCACGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 20 (RPI20) CAAGCAGAAGACGGCATACGAGATGGCCACGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 21 (RPI21) CAAGCAGAAGACGGCATACGAGATCGAAACGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 22 (RPI22) CAAGCAGAAGACGGCATACGAGATCGTACGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 23 (RPI23) CAAGCAGAAGACGGCATACGAGATCCACTCGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 24 (RPI24) CAAGCAGAAGACGGCATACGAGATGCTACCGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 25 (RPI25) CAAGCAGAAGACGGCATACGAGATATCAGTGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 26 (RPI26) CAAGCAGAAGACGGCATACGAGATGCTCATGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 27 (RPI27) CAAGCAGAAGACGGCATACGAGATAGGAATGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 28 (RPI28) CAAGCAGAAGACGGCATACGAGATCTTTTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 29 (RPI29) CAAGCAGAAGACGGCATACGAGATTAGTTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 30 (RPI30) CAAGCAGAAGACGGCATACGAGATCCGGTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 31 (RPI31) CAAGCAGAAGACGGCATACGAGATATCGTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 32 (RPI32) CAAGCAGAAGACGGCATACGAGATTGAGTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 33 (RPI33) CAAGCAGAAGACGGCATACGAGATCGCCTGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 34 (RPI34) CAAGCAGAAGACGGCATACGAGATGCCATGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 35 (RPI35) CAAGCAGAAGACGGCATACGAGATAAAATGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 36 (RPI36) CAAGCAGAAGACGGCATACGAGATTGTTGGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 37 (RPI37) CAAGCAGAAGACGGCATACGAGATATTCCGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 38 (RPI38) CAAGCAGAAGACGGCATACGAGATAGCTAGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 39 (RPI39) CAAGCAGAAGACGGCATACGAGATGTATAGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 40 (RPI40) CAAGCAGAAGACGGCATACGAGATTCTGAGGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 41 (RPI41) CAAGCAGAAGACGGCATACGAGATGTCGTCGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 42 (RPI42) CAAGCAGAAGACGGCATACGAGATCGATTAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 43 (RPI43) CAAGCAGAAGACGGCATACGAGATGCTGTAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 44 (RPI44) CAAGCAGAAGACGGCATACGAGATATTATAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 45 (RPI45) CAAGCAGAAGACGGCATACGAGATGAATGAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 46 (RPI46) CAAGCAGAAGACGGCATACGAGATTCGGGAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 47 (RPI47) CAAGCAGAAGACGGCATACGAGATCTTCGAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA RNA PCR Primer, Index 48 (RPI48) CAAGCAGAAGACGGCATACGAGATTGCCGAGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA Oligonucleotide sequences for Genomic DNA Adapters 5' P-GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT PCR Primers 5' AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT 5' CAAGCAGAAGACGGCATACGAGCTCTTCCGATCT Genomic DNA Sequencing Primer 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT Paired End DNA oligonucleotide sequences PE Adapters 5' P-GATCGGAAGAGCGGTTCAGCAGGAATGCCGAG 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT PE PCR Primer 1.0 5' AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT PE PCR Primer 2.0 5' CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT PE Read 1 Sequencing Primer 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT PE Read 2 Sequencing Primer 5' CGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT Oligonucleotide sequences for the Multiplexing Sample Prep Oligo Only Kit Multiplexing Adapters 5' P-GATCGGAAGAGCACACGTCT 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT Multiplexing PCR Primer 1.0 5' AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT Multiplexing PCR Primer 2.0 Multiplexing Read 1 Sequencing Primer 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT Multiplexing Index Read Sequencing Primer 5' GATCGGAAGAGCACACGTCTGAACTCCAGTCAC Multiplexing Read 2 Sequencing Primer 5' GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT PCR Primer, Index 1 CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTC PCR Primer, Index 2 CAAGCAGAAGACGGCATACGAGATACATCGGTGACTGGAGTTC PCR Primer, Index 3 CAAGCAGAAGACGGCATACGAGATGCCTAAGTGACTGGAGTTC PCR Primer, Index 4 CAAGCAGAAGACGGCATACGAGATTGGTCAGTGACTGGAGTTC PCR Primer, Index 5 CAAGCAGAAGACGGCATACGAGATCACTGTGTGACTGGAGTTC PCR Primer, Index 6 CAAGCAGAAGACGGCATACGAGATATTGGCGTGACTGGAGTTC PCR Primer, Index 7 CAAGCAGAAGACGGCATACGAGATGATCTGGTGACTGGAGTTC PCR Primer, Index 8 CAAGCAGAAGACGGCATACGAGATTCAAGTGTGACTGGAGTTC PCR Primer, Index 9 CAAGCAGAAGACGGCATACGAGATCTGATCGTGACTGGAGTTC PCR Primer, Index 10 CAAGCAGAAGACGGCATACGAGATAAGCTAGTGACTGGAGTTC PCR Primer, Index 11 CAAGCAGAAGACGGCATACGAGATGTAGCCGTGACTGGAGTTC PCR Primer, Index 12 CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGTTC