FIELD OF THE INVENTION
This invention relates to identification of cancer genes using a high-throughput forward genetic screening method called insertion mutation scanning (IMS).
BRIEF SUMMARY OF THE INVENTION
The invention includes a highly efficient and scalable method for large scale isolation of host/provirus fragments. The invention is an anchored-PCR method designed to amplify host/virus junction fragments from new clonally integrated proviruses present in tumors induced by certain retroviruses or pathogenic endogenous proviruses. These proviruses represent the somatic mutations which caused the tumors from which the integrants were isolated. The IMS technique has utility with respect to a cancer gene discovery strategy called provirus tagging (see, e.g., US 2004/0142336A1). Because of the efficiency and scalability of the IMS procedure, it is useful technology that makes a saturation screen of the cancer genome feasible. The IMS procedure implements an anchored-PCR approach to provirus tagging. Advantages over previous methods include improvements in (1) sensitivity, (2) signal-to-noise ratio, (3) efficiency of mutation recovery, and (4) scalability. The invention includes nucleotide sequences (both complete sequences and host sequences) shown in Table 1.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a schematic of bioinformatics analysis.
FIG. 2 shows the amplification results from 24 independent WXG/WLC tumors induced by MMTV. The upper common band contains the env region flanking the 3′ LTR and the lower common band contains the host sequence flanking the endogenous Mtv2 provirus which is common to all the tumors in the panel. Each unique band contains the 5′ host flanking sequence from a unique provirus insertion mutation that tags a cancer gene.
DETAILED DESCRIPTION
The methods of the invention can be used to prepare nucleotide sequences from cancer genes. A sample protocol illustrating the methods of the invention is as follows.
1. Extract genomic DNA from frozen tumor tissues using a SNAP-O-SOL kit from Biotecx (Houston, Tex.), according to the manufacturer's instructions.
2. Digest 750 ng of genomic DNA with 10 units of NlaIII from New England BioLabs (Beverly, Mass.) in a total of 20 μl for one hour at 37° C. in the manufacturer's buffer (NEBuffer 4 with supplemental BSA).
3. Heat inactivate the enzyme at 65° C. for 25 min and place onto ice.
4. Ligate anchors onto 150 ng of NlaIII digested DNA (4 μl). Anchors consist of two annealed oligos with a 3′ CATG overhang. The oligos, synthesized by Integrated DNA Technologies (Coralville, Iowa), have the following sequence:
- n0131 5′-CCTGA CTATG GAGTT CACTG CGTGC TACAC TATCA GTAGG CATG -3′
- n0157 5′-CCTAC TGATA GTGTA GCACG-N-3′
- note: n0157 contains a 3′ amino modifier (with a C7 spacer).
- Prior to use, dilute oligos from a concentrated stock solution to 4 μM each in TE.
- Mix equal parts to make a 2 μM solution.
- Heat to 95° C. for 2 min and lower temperature to 45° C. for 10 min.
5. Ligation:
DNA Ligation Kit Version 1 from Takara Shuzo Co., Ltd. (Otsu; Shiga, Japan)
- 4 μl NlaIII digested DNA
- 1 μl 2 μM n0131/n0157
- 25 μl Solution A
- 05 μl Solution B
- Mix all components on ice. Incubate at 16° C. for 1 hour.
- Purify using Qiaquick PCR Purification Kit from Qiagen, Inc. (Santa Clara, Calif.) according to the manufacturer's instructions. Elute into 50 μl 10 mM Tris-HCl, pH 8.5 and place on ice.
6. Amplify the ligated material through two successive rounds of PCR using nested primers as follows.
7. PCR reaction 1:
- 5 μl anchor ligated DNA
- 1X Qiagen PCR Buffer with 1.5 mM MgCl2 (Qiagen; Santa Clara, Calif.)
- 200 μM dNTPs
- 50 U/ml HotStarTaq DNA Polymerase (Qiagen; Santa Clara, Calif.)
- 0.2 U Perfect Match PCR Enhancer (Stratagene; LaJolla, Calif.)
- 0.2 μM of each of the amplification primers
- n0121 5′-CCTGA CTATG GAGTT CACTG-3′
- n0138 5′-GATGG GCTCA TCCGT TTGTG-3′
- H20 to a 50 μl reaction volume.
- Perform the reactions in an Amplitron II thermal cycler from Thermolyne (Dubuque, Iowa) using a step down PCR reaction with denaturation, annealing, and extension temperatures of 94° C., 60-56° C., and 72° C., respectively. During the first 5 cycles, drop the annealing temperature in one degree increments from 60° C. to 56° C. Perform the last 60 cycles at a constant annealing temperature of 56° C., for a total of 65 cycles during the first round.
- Cycling conditions:
- 95° C., 15 min dwell
- 2 cycles (94° C., 30 sec; 60° C., 30 sec; 72° C., 30 sec)
- 1 cycle (94° C., 30 sec; 59° C., 30 sec; 72° C., 30 sec)
- 1 cycle (94° C., 30 sec; 58° C., 30 sec; 72° C., 30 sec)
- 1 cycle (94° C., 30 sec; 57° C., 30 sec; 72° C., 30 sec)
- 60 cycles (94° C., 30 sec; 56° C., 30 sec; 72° C., 30 sec)
- 72° C., 10 min dwell
8. PCR reaction 2:
9. Shotgun clone 1 μl of this PCR product using a TA TOPO Cloning Kit from Invitrogen (Carlsbad, Calif.), according to the manufacturer's instructions with the following exception. Do not incubated transformed cells in a shaker after heat shock and subsequent ice step. Plate directly onto prewarmed LB Amp100/Kan 5 plates to ensure more accurate cloning representation.
10. PCR screen white colonies for insert size as described below.
11. Grown all screened colonies overnight in 2 ml LB Amp100/Kan50.
12. Miniprep cultures with correct insert sized using a QIAprep Miniprep Kit from Qiagen (Santa Clara, Calif.).
13. Send plasmids to University of California, Davis DBS Automated Sequencing Facility for insert sequence determination using PRISM sequencers, reagents, and software from ABI (Foster City, Calif.).
14. PCR screening for insert size:
- Pick 24 white colonies/tumor into 20 μl LB Amp100/Kan50.
- PCR reaction:
- 1 μl culture
- 1X Qiagen PCR Buffer with 1.5 mM MgCl2 (Santa Clara, Calif.)
- 200 μM dNTPs
- 20 U/ml Qiagen Taq Polymerase
- 0.2 μM each of the amplification primers
- n0122 5′-CGTGC TACAC TATCA GTAGG-3′
- n0036 5′-CAACC CCTTG GCTGC TTCTC-3′
- 50 μl reaction volume.
Perform the reactions in an Amplitron II thermal cycler from Thermolyne (Dubuque, Iowa) for 30 cycles with denaturation, annealing, and extension temperatures of 94° C., 56° C., and 72° C., respectively.
TABLE 1
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IST Sequences
INSERTCANCER
TAGBANDCOMPLETE SEQUENCEHOST SEQUENCELENGTHGENE
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IST000001MM0001BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATCATGAAGACCCCAGGCTCAGTT264unknown
GAAGACCCCAGGCTCAGTTTCTGCTCCCAGGAGAATCTGCTCCCAGGAGAAAGTCCT
AGTCCTGCTCTCTAGTCTGCCCTGAGGGCCAAACAGCTCTCTAGTCTGCCCTGAGGG
CGGGACTCTTGGTGCACCTCGTCAGTTTTGTGCAGCCAAACACGGGACTCTTGGTGC
TGCTTTAGCAGGGCCAGAGAGCCCATCTAGTGGCCACCTCGTCCAGTTTTGTGCAGT
AGGGCAGAACCCTGTCCCTGGCTGCATCTGGTCTTGCTTTAGCAGGGCCAGAAGCCC
CAGCTCAGGAATGTGAGATTGTCTTGTTAGTTTTTATCTAGTGGCCAGGGCAGAACC
GAAGTAACACAGTTCCTGCCTAGCTCCAAGAGTGGCTGTCCCTGGCTGCATCTGGTC
CGCTCCCTCCCACTTGCCGCGCCTGCAGCAGAAATTTCAGCTCAGGAATGTGAGATT
GGTTGAACTCCCGAGAGTGTCCTACACTTAGGGGAGTCTTGTTAGTTTTTGAAGTAA
GAAGCAGCCAAGGGGTTGAAGGGCGAATTCCAACAGTTCCTGCCTAGCTCCA
AGAGTGGCGCTCCTCCCACT{circumflex over ( )}
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IST000002MM0001CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATCATGCTCATAATTCTGCAGTGC156unknown
GCTCATAATTCTGCAGTGCCTTCTCATAACACAGGCTTCTCATAACACAGGATAAAA
ATAAAACACTCTAACCTTTAACATTATACTTGAAACACTCTAACCTTTAACATTATA
ACTTATGTGGTTTTTTCCTACCGAGTCATATCAAACTTGAAAACTTATGTGGTTTTT
CCAGTCTCCCTCTCCACTCACAAGGATCCAGTCACTCCTACCAGAGTCATATCAAAC
AATGGCCTTTTATGCCGCGCCTGCAGCAGAAATGGCAGTCTCCCTCTCCACTACAAG
TTGAACTCCCGAGAGTGTCCTACACTTAGGGGAGAGATCCAGTCACAATGGCCTTTT
AGCAGCCAAGGGTTGAAGGGCGAATTCA{circumflex over ( )}
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IST000003MM0001DGAATTCGCCCTCGTGCTACACTATCAGTAGGCATCATGTCCTACAGTGAACATTTC130unknown
GTCCTACAGTGAACATTTCTATATTTTCCACTTTTTATATTTTCCACTTTTTTCAGT
TTCAGTTTTCCTCGCCATATTTCACGTCCTAAAGTTTTCCTCGCCATATTTCACGTC
GTGTATTTCTCATTTTCCGTGATTTCAGTTTTCTCCTAAAGTGTGTATTTCTCATTT
GCCATATTCCAGGTCCTACAGTGCCGCGCCTGCAGTCCGTGATTTTCAGTTTTCTCG
CAGAAATGGTTGAACTCCCGAGAGTGTCCTACACTCCATATTCCAGGTCCTAAG{circumflex over ( )}
TAGGGGAGAAGCAGCCAAGGGGTTGAAGGGCGAAT
TC
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IST000004MM0001EGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATCATGGCGAGATTCTGTGTCCAA101Wnt3
GGCGAGATTCTGTGTCCAAGCTGCCTCTACTCGTGGCTGCCTCTACTCGTGACATTC
ACATTCCAAGATGCCTCTGAGGTGGGAACTGTGAACAAGATGCCTCTGAGGTGGGAA
ATAGGAGAGAGCCCCACAGTCCCTCTTTGCCGCGCCTGTGAAATAGGACAGAGCCCC
CTACAACAAAAATGGTTGAACTCCCGAGAGTGTCCACAGTCCCCTCTT{circumflex over ( )}
TACACTTAGGGGAGAAGCAGCCAAGGGGTTGAAGG
GCGAATTC
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IST000005MM0007CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATCATGTGTTTCCCGATTTTAGTT89unknown
GTGTTTCCCGATTTTAGTTGGATTTTTTTTCTCCCGGATTTTTTTTCTCCCAGGTTT
AGGTTTCTGCAGTGTCCCCACCCCCCACCCCCATTCTGCAGTGTCCCCACCCCCCAC
TTCTCCCACTCTTACCTGCCGCCCTGCAGCAGAAACCCCATTTTCTCCCACTCTTAC
TGGTTGAACTCCCGAGAGTGTCCTACACTTAGGGGC{circumflex over ( )}
AGAAGCAGCCAAGGGGTTGAAGGGCGAATTC
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IST000006MM0014BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATCATGGTGACAGTCCCTGAGTAA106unknown
GGTGACAGTCCCTGAGTAAAGGCTGGCTACTCCATAGGCTGGCTACTCCATCTTAGT
CTTAGTGGATTTGTTTTTATAAAGAAGGCAGACGAGGATTTGTTTTTATAAAGAAGG
GAACCAGAAATAACAACACATTTAAGCGTATGTGCCAGACGAGAACCAGAAATAACA
CGCGCCTGCAGCAGAAATGGTTGAACTCCCGAGAGACACATTGTAAGCGTATG{circumflex over ( )}
TGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTT
GAAGGGCGAATTC
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IST000007MM0008BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATCATGTATTTTTGCTGGGACTGT50unknown
GTATTTTTGCTGGGACTGTTCCCTCTGGGAACTGCTCCCTCTGGGAACTGCGGGATG
GGGATGGGGGTGTGCCGCGCCTGCAGCAGAAATGGGGGGTG{circumflex over ( )}
TTGAACTCCCGAGAGTGTCCTAACTTAGGGGAGAA
GCAGCCAAGGGGTTGAAGGGCGAATTC
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IST000008MM0003BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGGCAAGATGGAGACTTTG75Fgf3
GCAAGATGGAGACTTTGTCTACCAGGGCCACTCCAATCTACCAGGGCCACTCCAAGC
GCACCCAGCTGCATACAGGTGGACTGGCTGTGGCCTACCCAGCTGCATACAGGTGGA
GCCGCGCCTGCAGCAGAAAGGTTGAACTCCCGAGAGCTGGCTGTGGCC{circumflex over ( )}
TGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTTG
AAGGGCGAATTC
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IST000009MM0010BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGTAAGAAAAACAAGATCA170unknown
TAAGAAAAACAAGATCAAAGAAATGAGTCAAAAAGGAAGAAATGAGTCAAAAAGGCT
CTCTAACAGGGCTTGAAGGTGAGGTAGAAATGACAGCTAACAGGGCTTGAAGGTGAG
AAGACATAGAAATCCAATATCAGAAGGCAGAGTTTGGTAGAAATGACAGAAGACATA
TATGGGACAGATACTCTCTATATATATGCTAGCACAGAAATCCAATATTCAGAAGGC
CCTACTACTCAGCTGTTTCTCTGCCGCGCCTGCAGCAGAGTTTGTATGGGACAGATA
AGAAATGGTTGAACTCCCAAGAGTGTTCTACACTTACCTCTATATATATGCTAGCAC
GGGAGAAGCAGCCAAGGGGTTGAAGGGCGAATTCACCTACTACTCAGCTGTTTCT
C{circumflex over ( )}
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IST000010MM0015BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGTCCTACAGTGGATATTT135unknown
TCCTACAGTGGATATTTCTAAATTTTCCTCCTTTTTCTAAATTTTCCTCCTTTTTCA
CAGTTTTCCTCGCCATATTTGAAGTCCTAAAGTGTGGTTTTCCTCGCCATATTTGAA
TATTTCTCATATCCTGTGATTTCAGTTTTCTCSCCAGTCCTAAAGTGTGTATTTCTC
TATTCCAGGTCCTACAGTGTGCTGCCGC0CCTGCAGATATCCTGTGATTTTCAGTTT
CAGAAATGGTTGAACTCCCGAGAGTGTCCTACACTTTCTCGCCATATTCCAGGTCCT
AGGGGAGAAGCAGCCAAGGGGTCGAAGGGCGAATTCAAGTGTGC{circumflex over ( )}
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IST000011MM0015CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGTGGAGGCCAGAAGTCAA59unknown
TGGAGGCCAGAAGTCAACATATAGTCTCCTTCCCAACATATAGTCTCCTTCCCAATT
TTACTTGTCACTGGAGAGCTGCCGCGCCTGCAGCAGACTTGTCACTGGAGAGC{circumflex over ( )}
AAATGGTTGAACTCCCGAGGTGTCCTACACTTAGGG
GAGAAGCAGCCAAGGGGTTGAAGGGCGAATTC
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IST000012MM0013BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGCTACATTTTCATATTGT56unknown
CTACATTTTCATATTGTGTGCATACAAGGTTTCCCTGTGCATACAAGGTTTCCCTTT
TTATTATGTGGGGTAATGCCGCGCCTGCAGCAGAAAATTATGTGGGGTAA{circumflex over ( )}
TGGTTGAACTCCCGAGAGTTCCTACACTTAGGGGAG
AAGCAGCCAAGGGGTTGAAGGGCGAATTC
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IST000013MM0013BGAATTCGCCCTTCGTGCTACACTATCAGTAGGAATGAATGGGAGGAAGGACTGTGGA43unknown
GGAGGAAGGACTGTGGAAGGCAGAGGTGGAAATATCAGGCAGAGGTGGAAATATCTG
TGCTGCCGCGCCTGCAGCAGAAATGGTTGAACTCCCC{circumflex over ( )}
GAGAGTGTCCTACACTTAGGGAGAAGCAGCCAAGGG
GTTGAAGGGCGAATTC
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IST000014MM0006BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGCTGGCTGTTCCTGCAGC46Fgf8
CTGGCTGTTCCTGCAGCCCAGCTACTAGGACAATCTCCAGCTACTAGGACAATCTGG
GGAAACTGCCGCGCCTGCAGCAGAAATGGTTGAACTAAAC{circumflex over ( )}
CCCGAGAGTGTCCTACACTAGGGGAGAAGCAGCCAA
GGGGTTGAAGGGCGAATTC
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IST000015MM0007BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGGAAAGATAATGTGTAAA244unknown
GAAAGATAATGTGTAAATTTGGGTTTGCCGTGGAAATTTGGGTTTGCCGTGGAAAAC
ACTTTGGTTTCTCCATCAATGGTAATTGAGAGTTTGTTTGGTTTCTCCATCAATGGT
GCTGGGTATAGTAGCCTGGCTGGCATTTTTGTTCTCAATTGAGAGTTTGGCTGGGTA
TTAAGGTCTGTATAAAGTCTGTCCAGGATCTTCTGATAGTAGCCTGGGCTGGCATTT
CTTTCATAATGTCTGGTGTAAAGTCTGGTGTAATTCTTGTTCTCTTAAGGTCTGTAT
TGACAGGCCTGCCTTTATATGTTACTTGACCTTTTTAAGTCTGTCCAGGATCTTCTG
CCTTACTGCTTTTAATATTCTATGCCGCGCCTGCAGACTTTCATAATGTCTGGTGTA
CAGAAATGGTTGAACTCCCGAGAGTGTCCTACACTTAAGTCTGGTGTAATTCTGACA
AGGGGAGAAGCAGCCAAGGGGTTGAAGGGCGAATTCGGCCTGCCTTTATATGTTACT
TGACCTTTTTCCCTTACTGCT
TTTAATATTCTA{circumflex over ( )}
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IST000016MM0010CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCATGGGTCCCTGAAGGGTCTC79unknown
GGTCCCTGAAGGGTCTCTCCTTTAGCAAACCCCTGTTCCTTTAGCAAACCCCTGTAC
ACAGTTGAAGTGATTTTTCAGGTACCCATTGGTCTTAGTTGAAGTGATTTTTCAGGT
AGCTGCCGCGCCTGCAGCAAAATGGTTGAACTCCCGACCCATTGGTCTTAGC{circumflex over ( )}
AGAGTGTCCTACACTTAGGGGAGAAGCAGCCAAGGG
GTTGAAGGGCGAATTC
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IST000017MM0004BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCACATGTTGTCCTTTGAGCATCTC127unknown
TGTTGTCCTTTGAGCATCTCCACCTAGCAGCTGACACCTAGCAGCTGACTTAGACA
CTTAGACAGATACACACACCCACAGGTGAACAGTGATACACACACCCACAGGTGAA
GGATGGAGATTGGGGACTCTTATGGAGAATGGGACAGTGGATGGAGATTGGGGACT
GGAAGGATTTTGGGCCCTAAGGTGCCGCGCCTGCCTTATGGAAGAATGGGAGGAAG
AGCAGAAATGGTTGAACTCCCGAGAGTGTCCTACGATTTTGGGCCCTAAGG{circumflex over ( )}
ACTTANGGGAGAAGCAACCAAGGGGTGAAGGGCG
AATTC
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IST000018MM0004CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCACATGTGACACTGGCTTTGGAGT81unknown
TGTGACACTGGCTTTGGAGTCATACATAGAAGGGCATACATAGAAGGGGTTGGGAC
GTTGGGACAATTGATGCTAGTTAGCTAGAGCTAAAATTGATGCTAGTTAGCTAGAG
GAATATAGCAGTGCCGCGCCTGCAGAGAAATGGTCTAAGAATATAGCAG{circumflex over ( )}
TGAACTCCCGAGAGTGTCCTACACTTAGGGGAGA
AGCAGCCAAGGGGTTTAAGGGCGAATTC
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IST000019MM0005BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCACATGGTCTCTAGAGCTAAGAGA173unknown
TGGTCTCTAGAGCTAAGAGATACCAATGCTGCGGTACCAATGCTGCGGCAGGCAGT
CAGGCAGTTTTTATTACAATCATTACAGTTTTGATTTTATTACAATCATTACAGTT
CAGCGTCTGGCCGTGTGCCAAGGCTGCCTTCATCTTGACAGCGTCTGGCCGTGTGC
CCTGAGCTCGGTGATGCTTCTGTCCTGGTCTTCTCAAGGCTGGCCTTCATCCCTGA
GGCTCATCACAGCTTAAGAAAGTAGCTGCTTCTCGCTCGGTGATGCTTCTGCCTGG
TGCCGCGCCTGCAGCAGAAATGGTTGAACTCCCGTCTTCTGGCTCATCACAGCTTA
AGAGTGTCCTACACTTGGGGAGAAGCAGCCAAGGAGAAAGTAGCTGCTTCTC{circumflex over ( )}
GGTTGAAGGGCGAATTC
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IST000020MM0002CGAATTCGCCCTTCGTGCTACACTATCAGTAGGGAGATGGAACACTCAAAGCTGGCC115unknown
TGGAACACTCAAAGCTGGCCAGGGCCCATTTACCAGGGCCCATTTACCAGGTATCC
AGGTATCCTTTGCCTTCTCAGCTGATGGGCATCATTTGCCTTCTCAGCTGATGGGC
ACACATTAATTCACATATGACTCGTTGTGTCATAATCAACACATTAATTCACATAT
TCAATAGTATTGCCGCGCCTGCAGCAGAAATGGTGACTCGTTTGTGTCATATCAAT
TGAACTCCCGAGAGTGTCCTACACTTAGGGGAGAAGTAT{circumflex over ( )}
AGCAGCCAAGGGGTTGTTTCCCACCAAGGACGAC
CCGTCTGCGCACAAACGATGAGCCCATCAAGGGC
GAATTC
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IST000021MM0002BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCACATGGGGGGGGGTACCAAGAAG271unknown
TGGGGGGGGGTACCCAAGAAGGGACTGCTGCGATGGACTGCTGCGATTGGGATGTA
TGGGATGTAAATAAATAAATAAATAGAATAAACAAATAAATAAATAAATAGAATAA
AAACCCAAAAACAAACAGAAACCTAACTCAATAAACAAAACCCAAAAACAAACAGA
CTGCAGAAATGACTCTTGCTCTTTTCTGGTAAGGAACCTAAACTCAATAACTGCAG
TTAGAAGCAGGTTACAAATCTATATTAGAGATGGAAATGACTCTTGCTCTTTCTGG
AGGCATTTCACACCAGCATAGGTATAGGAAGTAGTAAGGTTAGAAGCAGGTTACAA
ATGAAATGAGGACTACCTAGAGTCTGTTTGTCACATCTATATTAGAGATGGAGGCA
AACCAATTCTGAGTGATTTCACTGAGATATTGCCTTTCACACCAGCATAGGTATAG
GCGCCTGCAGCAGAAATGGTTGAACTCCCGAGAGGAAGTAGATGAAATGAGGACTA
TGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTCACTAGAGTCTGTTTGTCACAA
TGAAGGGGAATTCCCAATTCTGAGTATTTCACTGA
GATAT{circumflex over ( )}
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IST000022MM0016DGAATTCGCCCTTCGTGCTACACTATCAGTAGGGAGATGTACTGAATCCCTGAAGTT66unknown
TGTACTGAATCCCTGAAGTTGATGCTGAGCACCAGATGCTGAGCACCATCTTGTGC
TCTTGTGCTGTTCTACCGCATTTACTGGGGTGCCTGTTCTACCGCATTTACTGGGG{circumflex over ( )}
GCGCCTGCAGCAGAAATGGTTGAACCCCGAGAGT
GTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTT
GAAGGGCGAATTC
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IST000023MM0016BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCACATGAGTACATTTCATTTTTTC212unknown
TGAGTACATTTCATTTTTTCCCACAAACTGCACACCACAAACTGCACAGACCAACT
GACCAACTTTGAGAAATGCCTGTTTATGAATGACTTGAGAAATGCCTGTTTATGAA
CAGAGATCACGTCTTCCATTTTTAACACTTAGAGTGACCAGAGATCACGTCTTCCA
ATGTGGAAGACTTCTGGTTAGCCCACTTATCCCTTTTTTAATCACTTAGAGATGTG
TCCCAGACTAGCAGTCAAGGCCAAGCAAAGGGAGGAAGACTTCTGGTTAGCCACTT
AATTTGGGCTGGTCCTATCCCCCCTGCTGGCTACATCCCTTCCCAGACTAGCAGTC
TGAACTGCCGCGCCTGAGCAGAAATGGTTGAACTAAGGCCAAGCAAAGGGAGAATT
CCCGAGGGTGTCCTACACTTAGGGGAGAAGCAGCTGGGCTGGTCCTATCCCCCCTG
CAAGGGTTGTTTCCCACCAAGGACGACCCGTCTGCTGGCTACTGAAC{circumflex over ( )}
CGCACAAACGGATGAGCCCATCAAGGGCGAATTC
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IST000024MM0016CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGATTATCTCTCTTCTATCCTT125unknown
CATGGATTATCTCTCTTCTATCCTTCCATTTTCCATTTTTGTTTTTGTTTGTTGGGT
TGTTTTTGTTTGTTGGGTTGGTTTTGATTTTTTGGTTTTGATTTTTGAGGCAGGGTT
GAGGCAGGGTTTCCCTGTGCATCCCTGCCTGCTCCCTGTGCATCCCTGCCTGGCTAT
TATCCTCGAACTTGACCTCACTCTGTAGCGCCCCTCGAACTTGACCTCACTCTGTAG{circumflex over ( )}
GCGCCTGCAGCAGAAATGGTTGAACTCCCGAG
AGTGTCCTACACTTAGGGGAGAAGCAGCCAAG
GGGTTGAAGGGCGAATTC
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IST000025MM0009CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTAATGCACAATATATGAGTCC96unknown
CATGTAATGCACAATATATGAGTCCTGATATTTGATATTTATCCCTTACAGTCAATA
TATCCCTTACAGTCAATATAGCTTTGTCTACATAGCTTTGTCTACAAATTTTTGACA
AATTTTTGACATAAAGTAGGGCTATTTTTCATTAAAGTAGGGCTATTTTTCAA{circumflex over ( )}
GCCGCGCCTGCAGCAGAAATGGTTGAACTCCC
GAGAGTGTCCTACACTTAGGGGAGAAGCAGCC
AAGGGGTTGAAGGGCGAATTC
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IST000026MM0009BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGAAATGCAATGAAAGCACACA301Unknown
CATGGAAATGCAATGAAAGCACACAATATTGTATATTGTTGAACCAAACAGAAAGCT
TGAACCAAACAGAAAGCTCAAAATTAGGCACACAAAACTAGGCACAGAAAAGAGATA
GAAAAGAGATACAAACACAAATCTGAACAAATCAAACACAAATCTGAACAAATTGAC
GACCTTCTCCCTATAGCATAACTAATATCTCACTTCTCCCTATAGCATAACTAATAT
GAGATAAAAGTGGTCTTTATATACCAGGGCGACTAGAGATAAAAGTGGTCTTTATAT
AAGAGGTCTAAAAAGAGAGAAATAAAAAATATACCAGGGCGAAAGAGGTCTAAAAAG
GGCATATTTCCTGTCATATGCAGAACCTATTGAGAGAAATAAAAAATATGGCATATT
AGTCTTTTTGTTTGTTTCTTTCAATACAGCCTTCCTGTCATATGCAGAACCTATATG
ATGTAGCTCTAGCTGTCCTAGAACTTACTTTGAGTCTTTTTGTTTGTTTCTTTCAAT
TAGACCAGGCTTGCCGCGCCTGCAGCAGAAATACAGCTATGTAGCTCTAGCTGTCCT
GGTTGAACTCCCGAGAGTGTCCTACACTTGGGAGAACTTACTTTGTAGACCAGGCT{circumflex over ( )}
GAGAAGCAGCCAAGGGGTTGAAGGGCGAATTC
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IST000027MM0011BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTTGACAGTCCTAGAGCAAGAC163unknown
CATGTTGACAGTCCTAGAGCAAGACCTGAGGTCTGAGGTAGAAGAATGGCCTTTAGA
AGAAGAATGGCCTTTAGAAAGTAGGGACTTCAAAGTAGGGACTTCAGGTGCCTGAAA
GGTGCCTGAAAAATGGAGGGGTGCCAGGGCAAAATGGAGGGGTGCCAGGGCAGATGT
TGTGGGGAGCCCTGTGAGGAGTGGGGTGTCCCGGGGAGCCCTGTGAGGAGTGGGGTG
AGGGCAGATGTGGGGAGCCCTGTGAGGAGTGGTCCAGGGCAGATGTGGGGAGCCCTG
GGTGCCGCGCCTGCAGCAGAAATGGTTGAACTTGAGGAGTGGGG{circumflex over ( )}
CCCGAGAGTGTCCTACACTTAGGGGAGAAGAG
CCAAGGGGTTGAAGGGCGAATTC
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IST000028MM0011CGAATTCGCCCTTCGTGCTACACTATCAGTAGGGATGTCAGAGCCGCAGGAATGCATA90unknown
GATGTCAGAGCCGCAGGAATGCATAATGGTCTATGGTCTTCTTTTTGTTTGTTTGTT
TCTTTTTGTTTGTTTGTTTTTTGAGACAGGGTTTTTGAGACAGGGTTTCTCTGTGTA
TTCTCTGTGTAGCCCTGGCTGTCCTGTGCCGGGCCCTGGCTGTCCTG{circumflex over ( )}
CCTGCAGCAGAAATGGTTGAACTCCCGAGAGT
GTCCTACACTTAGGGGAGAAGCAGCCAAGGGG
TTGAAGGGCGAATTC
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IST000029MM0011DGAATTCGCCCTTGGTGCTACACTATCAGTAGGCATGCAGGAGGCCCTCTGTCCTTAA40unknown
CATGCAGGAGGCCCTCTGTCCTTAACACAGAACACAGAATGTACAGG{circumflex over ( )}
TGTACAGGTGCCGCGCCTGCAGCAGAAATGGT
TGAACTCCCGAGGGTGTCCTACACTTAGGGGG
AAGCAGCCAAGGGGTTGTTTCCCACCAAGGAC
GACCCGTCTGCGCACAAACGGATGAGCCCATC
AAGGGCGAATTC
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IST000030MM0010DGAATTCGCCCTTCGTGCTACACTATCAGTAGGGATGCCCGATCAGTTGCTTCTAGAC48unknown
GATGCCCGATCAGTTGCTTCTAGACATAAAGAATAAAGAAAAATTCGTTGTTTTC{circumflex over ( )}
AAAATTCGTTGTTTTCTGCCGCGCCTGCAGCA
GAAATGGTTGAACTCCCGAGAGTGTCCTACAT
TAGGGGAGAAGCAGCCAAGGGGTTGAAGGGCG
AATTC
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IST000031MM0012BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTGCATGTGAGTTCTTTTGTG187unknown
AGTTCTTTTGTGATTGGGTTACCTCACTCAGGATGATAATTGGGTTACCTCACTCA
CCCTCCAGATCCATCCATTTGCCTAGGAATGTCATAAAGGATGATACCCTCCAGAT
TTCATTCTTTTTATAGCTGAGTAGTACTCCATTGTGTACCATCCATTTGCCTAGGA
AATGTACCACACTTTCTGTATCCATTCCTCTGTTGAGGATGTCATAAATTCATTCT
GGCATCTGGGTTCTTTCCAGCTTCTGGCTGCCGCGCCTTTTTAATAGCTGAGTAGT
GCAGCAGAAATGGTTGAACTCCCGAGGTGTCCTACACTACTCCATTGTGTAAATGT
TAGGGGAGAAGCAGCCAAGGGGTAGAAGGGCGAATTCACACACTTTCTGTATCCA
TTCCTCTGTTGAGGGGCA
TCTGGGTTCTTTCCAGCT
TCTGGC{circumflex over ( )}
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IST000032MM0014DGAATTCGCCCTTCGTGCTACACTATCAGTAGGGATGATGATGATG{circumflex over ( )}7too short
GTGCCGCGCCTGCAGCAGAAATGGTTGAACTCCCGAGA
GTGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTTGA
AGGGCGAATTC
|
IST000033MM0018BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGACATGGATCCTGACAACAG223unknown
TCCTGACAACAGGCTGAGTAAATGAAGCTGTTTATTCTGCTGAGTAAATGAAGCTG
CAAGTGCTGAGGGCTGACCTGGAGCATACTGTCTAGAGTTTATTCTCAAGTGCTGA
GGACCACTGCAGTTTCAAGTATGGATGGTGTTGAGCTTGGGCTGACCTGGAGCATA
TGGGGGATGCTAGATGCATCTATAAATGTGTGCTGTTTCTGTCTAGAGGGACCACT
ACTTTTTTTCATCTTGGTATATAATCTTTATTTGTACTGCAGTCTTCAAGTATGGA
TTGTCAGTTTCATCTGTGTAGAAGATGCCGCGCCTGCATGGTGTTGAGCTTTGGGG
GCAGAAATGGTTGAACTCCCGAGAGTGTCCTACACTTAGTGCTAGATGCATCTATA
GGGGAGAAGCAGCCAAGGGGTTGTTTCCCACCAAGGACAATGTGTGCTGTTTACTT
GACCCGTCTGCGCACAAACGGATGAGCCCATCAAGGGCTTTTTCATCTTGGTATAT
GATTCAATCTTTATTTGTACTTT
GTCAGTTTCATCTGTGTA
GAAGAT{circumflex over ( )}
|
IST000034MM0018CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTTCATGTTCCTTTGTCAAGG100unknown
CCTTTGTCAAGGTCTATGGACATTTCTGTCTTTTGGAATCTATGGACATTTCTGTC
TGTTCCTCCTTTAGATATCCTGGGGTTTTGTTGTTGTTTTTTGGAATGTTCCTCCT
GTTTTGTTTTGAGCAGTTGCCGCGCCTGCAGCAGAAATTTAGATATCCTGGGGTTT
GGTTGAACTCCCGAGAGTGTCCTACACTTAGGGGAGAATGTTGTTGTTGTTTTGTT
GCAGCCAAGGGGTTGTTTCCCACCAAGGACGACCCGTCTTGAGGCAGT{circumflex over ( )}
TGCGCACAAACGGATGAGCCCATCAAGGCGAATTC
|
IST000035MM0019BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGAGCATGAGCTGTCGATAGTG129unknown
CTGTCGATAGTGACCTGCAGTCAAGGAAATCTGAGGGCACCTGCAGTCAAGGAAAT
TTCCTAATTAACAGAGGAGCTCTAAATGAGAGTAACGCCTGAGGGCTTCCTAATTA
GCTCCACAAACCCCTCACACTCGGTAAGTGTCACGGTGACAGAGGAGCTCTAAATG
CAGATAATTGCCGCGCCTGCAGCAGAAATGGTTGAACTAGAGTAACGCGCTCCACA
CCCGAGAGTGTCCTACACTTAGGGGAGAAGCAGCCAAGAACCCCCTCACACTCGGT
GGGTTGTTTCTCACCAAGGACGACCCTCTGCGCACAAAAAGTGTCACGGTGCAGAT
CGGATGAGCCCATCAAGGGCGAATTCAT{circumflex over ( )}
|
IST000036MM0021BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGAGCATGAGCACTAAAGGCAG105unknown
CACTAAAGGCAGCTAACCAAAACAGAATCCCACAGACTCTAACCAAAACAGAATCC
ACTCTGCAGCTTCTTAGGCTGGCTCGTTTTTCCAGTCTCACAGACTACTCTGCAGC
AGCTATTGCCTTATTCCTCTTCTGCCGCGCCTACAACATTCTTAGGCTGGCTCGTT
GAAATGGTTGAACTCCCGAGAGTGTCCTACACTTAGGGTTTCCAGTCTAGCTATTG
GAGAAGCAGCCAAGGGGTTGTTTCCCACCAAGGACGACCCTTATTTCCTCTTC{circumflex over ( )}
CCGTCTGCGCACAAACGGATGAGCCCTCAAGGGCGAAT
TC
|
IST000037MM0021CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGACATGGATAGAGGATGGAA66unknown
TAGAGGATGGAAGTTGAAAACCTGCTATTAAGAATATAGTTGAAAACCTGCTATTA
GCCCTGTCCATTAGTGAGAGTGTGCCGCGCCTGCAGCAAGAATATAGCCCTGTCCA
GAAATGGTTGAACCCCGAGAGTGTCCTACACTTAGGGGTTAGTGAGAGTG{circumflex over ( )}
AGAAGCAGCCAAGGGGTTGTTTCCCACCAAGGACGACC
CGTCTGCGCACAAACGGATGAGCCCATCAAGGGCGAAT
TC
|
IST000038MM0024BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGAGGGCATGAGGGGTACAAATT78unknown
GTACAAATTTGTTGTGCCTAAAAGCATTGAAGTGTAGGGATGTTGTGCCTAAAAGCA
AGACTGGGAAGTGGCACGGTCAGACTTAGGTGCCGCGCCTTTGAAGTGTAGGGAAGA
GCAGCAGAATGGTTGAACTCCCGAGAGTGTCCTACACTTACTGGGAAGTGGCACGGT
GGGGAGAAACAGCCAAGGGGTTGTTTCCCACCAAGGACGACAGACTTAGG{circumflex over ( )}
CCCGTCTGCGCACAAACGGATGAGCCCATCAAGGGCGAAT
TC
|
IST000039MM0024CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTCACCATGTCACACTGGGGAC46unknown
ACTGGGGACTCCCTCCGGGGAGGGAGCAGGCTGGGGGGTGTCCCTCCGGGGAGGGAG
CCGCGCCTGCAGCAGAAATGGTTGAACTCCCGAGAGTGTCCAGGCTGGGGGG{circumflex over ( )}
CTACACTAGGGGAGAAGCAGCCGAGGGGTTGTTTCCCACC
AAGGACGACCCGTCTGCGCACAAACGGATGAGCCCATCAA
GGGCGAATTC
|
IST000040MM0022BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGCTACATGGCTATCCTTTCCT99Wnt1
TCCTTTCCTTTCATTGCTGCTACCTCCCCAGCTCTTGCACTTCATTGCTGCTACCTC
TAGGAGAGCTGGACTGTTGGTGTGCTGAGTCCCCATCAGGCCCAGCTCTTGCACTAG
TGACACATTGTGCCGCGCCTGCAGCAGAAATGGTTGAACTGAGAGCTGGACTGTTGG
CCCGAGAGTGTCCTACACTTAGGGGAGAAGCAGCCAAGGGTGTGCTGAGTCCCCATC
GTTGTTTCCCACCAAGGACGACCCGTCTGCGCACAAACGGAGGTGACACACTTG{circumflex over ( )}
ATGAGCCCATCAAGGCGAATTC
|
IST000041MM0022CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTCAGCATGTCAGGTGGCTCAC85unknown
GTGGCTCACCTGCAGAGGATCTAATTAATGCCTCCACCCTCTGCAGAGGATCTAATT
CCAGGAGTGCCTACCCTCACATAAACATATCCATATATGCAATGCCTCCACCCTCCA
CGCGCCTCAGCAGAAATGGTTGAACTCCCGAGAGTGTCCTGGAGTGCCTACCCTCAC
ACACTTAGGGGAGAAGCAGCCAAGGGGTTGTTTCCCACCAATAAACATATCCATATA{circumflex over ( )}
AGGACGACCCGTCTGCGCACAAACGGATGAGCCCATCAAG
GGCGAATTC
|
IST000042MM0022DGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTCTGCATGTCTGCTGTCTAAA72unknown
CTGTCTAAACCTCCTAATCTATATATTTTGTTACAGATGTCCTCCTAATCTATATAT
TCAAGATGACTACAGTCCACATATTGCCGCGCCTGCAGCATTTGTTACAGATGTTCA
GAAATGGTGAACTCCCGAGAGTGTCCTACACTTAGGGGAGAGATGACTACAGTCCAC
AAGCAGCCAAGGGGTTGTTTCCCACCAAGGACGACCCGTCATAT{circumflex over ( )}
TGCGCACAAACGGATGAGCCCATCAAGGGCGAATTC
|
IST000043MM0022FGAATTCGCCCTTCGTGGTACACTATCAGTAGGCATGGTTCCATGGTTCTGGGAA{circumflex over ( )}14too short
TGGGAATGCCGCGCCTGCAGCAGAAATGGTTGAACTCCCG
AGAGTGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTTG
TTTCCCACAAGGACGACCCGTCTGCGCACAAACGGATGAG
CCCATCAAGGGCGAATTC
|
IST000044MM0022GGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTGCCCATG{circumflex over ( )}4too short
GCGCCTGCAGCAGAAATGGTTGAACTCCCGAGAGTGTCCT
ACACTTAGGGGAGAAGCAGCCAAGGGGTTGTTTCCCACCA
AGGACGACCGTCTGCGCACAAACGGATGAGCCCATCAAGG
GCGAATTC
|
IST000045MM0017BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTGTCCATGTGTCACTCAAAGG205unknown
ACTCAAAGGCTGCTGAGAATCAGGCTGTACCTGCATTCCTCTGCTGAGAATCAGGCT
AAGCCATCCACAGCCATCCTGACCCACAGCAAATGCTGGCGTACCTGCATTCCTAAG
AGTCGCCCACAGCTGGACTCCGTTCCTCCCTCCACTCCTACCATCCACAGCCATCCT
TAGCCGAGGCTATCCACACAGGCTATTTCAGTGCCCTAAGGACCCACAGCAAATGCT
CCTTGCTACCCTTATGTATACATTGAGGACAATGACTGCCGGCAGTCGCCCCACAGC
GCGCCTGCAGCAGAATGGTTGAACTCCCGAGAGTGTCCTATGGACTCCGTTCCTCCC
CACTAAGGGGAGAAGCAGCCAAGGGGTTGTTTCCCACCAATCCACTCCATAGCCGAG
GGACGACCCGTCTGCGCACAAACGGATGAGCCCATCAAGGGCTATCCACACAGGCTA
GCGAATTCTTTCAGTGCCCTAAGCC
TTGCTACCCTTATGTAT
ACATTGAGGACAATGAC{circumflex over ( )}
|
IST000046MM0017DGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGAACATGGAAATA{circumflex over ( )}10too short
ATATGCCCCGCCTGCAGCAGAAATGGTTGAACTCCCGAG
AGTGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTTGT
TTCCCACCAAGACGACCCGTCTGCGCACAAACGGATGAG
CCCATCAAGGGCGAA~TTC
|
IST000047MM0017EGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGCCGCA{circumflex over ( )}2too short
CGCCTGCAGCAGAAATGGTTGAACTCCCGAGAGTGTCCT
ACACTTAGGGGAGAAGCAGCCAAGGGGTTGTTTCCCACC
AAGGACGACCGTCTGCGCACAAACGGATGAGCCCATCAA
GGGCGAATTC
|
IST000048MM0020BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGAGCATGGAGAAACCTGGGCT126unknown
AAACCTGGGCTTATTCAAGCAGTTTCCTTTGTTTACCCTTATTCAAGCAGTTTCCTT
GCCCAGGGTTGCCAGTGAAGGGGCTCCTCCATCACTAACTGTTTACCCTGCCCAGGG
TAAAGGTCTTTCCTATGCTGGTTCCTCTCCACCCCACCATTGCCAGTGAAGGGGCTC
TTGCCGCGCCTGCAGCAGAAATGGTTGAACTCCCGAGAGCTCCATCACTAACTAAAG
TGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTTGTTTGTCTTATCCTATGCTGGT
CCCACCAAGGACGACCCGTCGCGCACAAACGGATGAGCCTCCTCTCCACCCCACCAT{circumflex over ( )}
CATCAAGGGCGAATTC
|
IST000049MM0020DGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGGGTCATGGGTGTG{circumflex over ( )}10too short
GTGTGCCGCGCCTGCAGCAGAAATGGCTGAACTCCCGAG
AGTGTCCTACACTTAGGGGAGAAGCAGCCAAGGGGTTGT
TTCCCACCAAGACGACCCGTCTGCGCACAAACGGATGAG
CCCATCAAGGGCGAATTC
|
IST000050MM0023BGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGTCCCATGTCCTCCACAGCCGA221Fgf3
TCCACAGCCGAGCACCCTCAACTGTCTCCCAGGGCCTCTGCACCCTCAACTGTCTCC
GTTCTAGCCAGGGTCTGCAGGGTCTCTGCCCCACGCCTACAGGGCCTCTGTTCTAGC
GCCCCTGAGAATCTTAAGCGGTCTGAAAACTACGCCACTCAGGGTCTGCAGGGTCTC
GAACTGCTAAAACCCTGGAGTCACTGATGGAACCTGCTTTGCCCCACGCCTAGCCCC
TACAGGGGACAGCCCTGTGTCTCGAAGACCAACCTAAGCTGAGAAATCTTAAGCGGT
TCCTAAGATGTCACAAATTGCGCGCCTGCAGCAGAAATGCTGAAAACTACGCCACTG
GTTGAACTCCCGAGAGTGTCCTACACTTAGGGGAGAAGCACTGCTAAAACCCTGGAG
AGCCAAGGGGTTGTTTCCCACCAAGGACGACCCGTCTGCTCACTGATGGAACCTGCT
GCACAAACGGATGAGCCCATCAAGGGCGAATCTTACAGGGGACAGCCCTG
TGTCTCGAAGACCAACCT
AAGGTCCTAAGATGTCAC
AAAT{circumflex over ( )}
|
IST000051MM0023CGAATTCGCCCTTCGTGCTACACTATCAGTAGGCATGACGCATGACGACTTGAAAAAT39unknown
ACTTGAAAAATGACGAAATCACTAAAAAACAATGCCGCGGACGAAATCACTAAAAAA
CCTGCAGCAGAAATGGTTGAACTCCCGAGAGTGTCCTACCAA{circumflex over ( )}
ACTTAGGGGAAAGCAGCCAAGGGGTTGTTTCCCACCAAG
GACGACCCGTCTGCGCACAAACGGATGAGCCCATCAAGG
GCGAATTC
|
It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.