TGXAS-甘蔗品种SNP分子检测技术规程

Q/LB.□XXXXX-XXXX前言本文件按照GB/T1.1—2020《标准化工作导则第1部分：标准化文件的结构和起草规则》的规定起草。请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别专利的责任。本文件由福建农林大学、广西大学提出并宣贯。本文件由广西标准化协会归口。本文件起草单位：福建农林大学、广西大学、广东省科学院南繁种业研究所。本文件主要起草人：赵新旺、张木清、邓祖湖、暴怡雪、赵薇彤、黄江锋、王勤南、姚伟、徐良年、吴嘉云、胡琴、张南南、张垂明、余凡、蒋洪涛、肖胜华、张伟。FORMTEXT甘蔗品种SNP分子检测技术规程范围本文件规定了构建甘蔗(Saccharumspp.hybrid)品种SNP分子标记指纹图谱的原理、检测方案、检测程序和结果判定。本文件适用于构建甘蔗常规品种的SNP分子标记指纹图谱和身份鉴定。规范性引用文件下列文件中的内容通过文中的规范性引用而构成本文件必不可少的条款。其中，注日期的引用文件，仅该日期对应的版本适用于本文件；不注日期的引用文件，其最新版本（包括所有的修改单）适用于本文件。GB/T3543.2农作物种子检验规程扦样GB/T3543.5农作物种子检验规程真实性和品种纯度鉴定GB/T6682分析实验室用水规格和试验方法GB/T19495.3转基因产品检测核酸提取纯化方法术语和定义下列术语和定义适用于本文件。品种真实性验证varietyverification与其对应品种名称的标准样品比较，检测并证实供检样品品种名称与标注是否与标准品种样品相符。多重PCRmultiplexPolymeraseChainReaction，mPCR在同一PCR反应体系里加上二对以上引物，同时扩增出多个核酸片段的PCR反应，其反应原理，反应试剂和操作过程与一般PCR相同。

连接酶检测反应LigaseDetectionReaction，LDR在TaqDNA连接酶的作用下，当左右两条寡核苷酸探针与目的DNA序列完全互补，并且两条探针之间没有空隙时才能发生连接反应，最后通过荧光扫描片段长度，实现对SNP位点的检测。缩略语下列缩略语适用于本文件。bp：basepair碱基对CTAB：cetyltrimethylammoniumbromide十六烷基三甲基溴化铵DNA：deoxyribonucleicacid脱氧核糖核酸PCR：polymerasechainreaction聚合酶链式反应LDR：ligasedetectionreaction连接酶检测反应SNP：singlenucleotidepolymorphisms单核苷酸多态性仪器设备及试剂仪器设备及试剂见附录A，所用试剂均为分析纯。试剂配制所用水应符合GB/T6682规定的一级水的要求。溶液配制溶液配制见附录B。SNP标记一套甘蔗品种真实性检测SNP标记包括177个核心SNP标记、位点及物理位置见附录C。引物信息引物分为多重PCR引物、连接引物见附录D。检测程序样品准备采集甘蔗叶片样品重量应不低于3g，并用自封袋包装，冰盒携带，4℃条件下保存。用混合样品检测的，每个个体上等量取样，混合后提取DNA。采集种茎的，常温运送，温室种植，待长出真叶后采集叶片取样，方法同上。采样脱毒组培瓶苗的，在提取DNA前不开瓶，防止污染。样品在4℃下可以保存2周，在-20℃下可以保存6个月，在-80℃下可以长期保存。设标准样品，与样品采集同样方法。从供检样品中分别取出有代表性、符合GB/T3543.2规定的试样，采用混合样或单个个体进行检测试样来源应至少含有3个个体。DNA样品准备参照GB/T19495.3的方法或者使用试剂盒进行DNA提取。保证提取的DNA数量和质量符合PCR扩增的要求，DNA无降解，溶液的紫外光吸光度OD260与OD280的比值介于1.8~2.0，经过琼脂糖电泳检测具有清晰明亮的单一主带。多重PCR反应将配制好的PCR反应预混合液充分震荡离心混匀后分装到96孔板里，每个反应孔分装19µL（表1）。然后每个反应孔里加入1µL的DNA样本，盖上盖膜，轻微震荡混匀后3,000rpm离心30秒，然后立即将离心后96孔板放置PCR仪上，去除盖膜，然后盖上96孔橡胶盖垫，盖好PCR仪热盖，按表2程序运行。177个甘蔗核心SNP标记分为4个多重LDR体系进行检测，见附录E。多重PCR反应预混合液配制试剂表试剂1×100×10×PerfectStart®Taqbuffer2µL200µLdNTP（2.5mM）2.4µL240µLMgCl2（25mM）0.8µL80µLProbeMix2.0µL200µLHotStarTaq（2.5U/µL）0.4µL40µLddH2O11.4µL1140µL总体积19µL1900µL多重PCR反应程序变性退火延伸保持循环数1step95ºC2min12step94ºC20sec65ºC40sec(-0.5ºC/cycle)72ºC1.5min-113step94ºC20sec59ºC30sec72ºC1.5min-244step--72ºC2.0min4ºC1连接反应将配制好的PCR反应预混合液充分震荡离心混匀后分装到96孔板里，每个反应孔分装6μL（表3）。每个反应孔里加入4μL的多重PCR纯化产物，盖上盖膜，轻微震荡混匀后3,000rpm离心30秒，然后立即将离心后96孔板放置PCR仪上，去除盖膜，然后盖上96孔橡胶盖垫，盖好PCR仪热盖，按表4程序运行。连接反应预混合液配制试剂表试剂1×100×10×ligasebuffer1.00μL100μLLigasePrimerMix(1μM)0.4μL40μLDNAligase(50U/μL)1.0μL100μLddH2O3.6μL360μL总体积6μL600μL连接反应程序变性连接保持循环数1step94ºC1.0min56ºC4.0min-382step--4ºC-上机检测连接产物加入到70μLddH2O，充分震荡离心后，取3μL与0.5μLLiz500SizeStandard，8.5μLHi-Di混匀，95ºC变性5分钟后用ABI3730XL测序仪分析检测。数据分析使用软件GeneMapper4.1（AppliedBiosystems,USA）进行数据分析。基因型判断每个位点的两个等位基因size大小不同，size大小在设计体系探针时已设计好，所以等位基因在峰图中的位置是固定的，根据峰型图即可判断出该位点的基因型。数据记录方式为具体基因型，数据记录为X/Y，其中X，Y为同一位点上两个不同的等位变异，即纯合基因型记录为XX或YY，杂合基因型记录为XY，缺失位点记录为NN。单位点基因型判断的峰图见附录F。真实性验证根据供检样品与对照样品在177个核心标记中的位点差异，进行品种真实性验证。将检测得到的基因型根据标记顺序进行排列，并计算基因型序列的遗传距离或遗传相似性。遗传距离小于0.02或遗传相似性大于98%的认定为同一品种，遗传距离大于等于0.02或遗传相似性小于等于98%的认定为不同品种。——当差异位点数大于等于5个时，认定为不同品种；——当差异位点数大于1且小于等于4时，认定为相似品种；——当差异位点数小于等于1时，认定为同一品种。

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主要仪器设备与试剂主要仪器设备PCR仪、全自动遗传分析仪、微量移液枪（2.5µL、10µL、100µL、1000µL）、电泳槽及配套的制胶配件、恒压电泳仪、凝胶成像系统、恒温水浴锅、制冰机、电子天平、酸度计、高压灭菌锅、高速台式离心机、冰箱、超净工作台、紫外分光光度计、酸浓度测定仪。主要试剂十六烷基三乙基溴化铵（CTAB）、三氯甲烷、异丙醇、异戊醇、乙二胺四乙酸二钠、三羟甲基氨基甲烷、盐酸、氢氧化钠、氯化钠、β-巯基乙醇（β-Mercaptoethanol）、dNTP混合液、Ex-TaqDNA聚合酶、10×Ex-TaqBuffer:含Mg2-25mMol/L、琼脂糖、DNA分子量标准、无水乙醇、SNP引物及连接引物、去离子水、DNA分析仪专用甲酰胺、DNA分析仪专用分子量内标Liz标记、DNA分析仪专用电泳缓冲液。甘蔗黑穗病菌冬孢子活性检测形态

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溶液配制0.5mol/LEDTA溶液称取186.1gNa2EDTA·2H2O溶于800mL水中，加固体NaOH调pH至8.0，加水定容至1,000mL，121℃高压灭菌20min。1mol/LTris-HCI溶液称取60.55gTris碱溶于适量水中，加HCI调制调pH至8.0，加水定容至500mL，121℃高压灭菌20min。5mol/LNaCl溶液称取146.1g固体NaCl，加水定容至500mL，121℃高压灭菌20min。CTAB提取液称取280mL的5mol/LNaCl、20mLβ-巯基乙醇（C2H6OS)、20gCTAB、100mL1mol/LTris-HCl和40mL0.5mol/LEDTA，混合加水定容至1000mL，4℃贮存。SDS提取液称取100mL1mol/LTris-HCl、25mL0.5mol/LEDT、25mL5mol/LNaCl和2.5gSDS，混合加水定容至500mL。5mol/LKAC称取49.67gKAC，加水溶解，冰乙酸调整pH至5.2，定容至100mL，121℃高压灭菌20min。1×TAE溶液称取5mL1mol/LTris-HCl和1mL0.5mol/LEDTA，加HCl调pH至8.0，加水定容至500mL，121℃高压灭菌20min。75%乙醇称取75mL无水乙醇和25mL蒸馏水混匀。

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甘蔗黑穗病抗性标准对照品种核心SNP标记位点及物理位置见表C.1。核心SNP标记位点及物理位置标记物序号SNP序号染色体等位基因物理位置1SNP001Chr01AdG/T989274872SNP002Chr01BdG/A154277953SNP004Chr01BdC/G832846284SNP006Chr01CdT/C290263035SNP008Chr01CdT/G605324616SNP010Chr01DdC/T140367327SNP011Chr01DdG/A271815688SNP014Chr01EdG/T499063969SNP015Chr01EdA/G9290680810SNP019Chr01FdT/A5297057711SNP023Chr01GdC/T6522980312SNP025Chr01GdG/A10363146813SNP028Chr01HdC/T5362985414SNP031Chr02AdT/A6760499615SNP032Chr02AdG/T10940227516SNP034Chr02BdG/A2705103817SNP036Chr02BdG/A10910715918SNP037Chr02CdG/A1304068419SNP039Chr02CdG/A4036303520SNP040Chr02CdG/T5602109921SNP042Chr02CdG/A9321589522SNP043Chr02DdT/G1400291923SNP045Chr02DdT/C6472684924SNP046Chr02DdC/T8968381225SNP048Chr02EdC/G2565640626SNP049Chr02EdT/C5096746727SNP050Chr02EdA/T6162575128SNP052Chr02FdG/A4799619729SNP053Chr02FdT/A7252741430SNP055Chr02FdG/A9361234931SNP059Chr02GdC/T4935742932SNP060Chr02HdC/T1977093233SNP062Chr03AdC/T4689901234SNP063Chr03AdA/T7024636835SNP065Chr03AdA/T9239948036SNP066Chr03BdC/A949724337SNP070Chr03BdC/T6811955038SNP071Chr03BdC/A8872315939SNP076Chr03DdG/A765025540SNP077Chr03DdC/T3210352441SNP079Chr03DdT/C7227097542SNP082Chr03EdG/A6031656443SNP084Chr03FdC/T2666824144SNP085Chr03FdC/T3822577745SNP086Chr03FdG/A4686651046SNP087Chr03FdC/T8103228547SNP088Chr03GdC/T1008060648SNP089Chr03GdC/T4809029149SNP090Chr03GdT/A5579925050SNP091Chr03GdA/G7635431851SNP092Chr03HdC/A1554846352SNP095Chr03HdC/A3081877053SNP096Chr03HdG/A5141006754SNP100Chr04BdG/C1053471055SNP101Chr04BdC/T2199943756SNP102Chr04BdC/T4509485457SNP103Chr04BdC/A5358584358SNP104Chr04CdA/T1102307659SNP107Chr04CdC/G6326331460SNP110Chr04DdG/A2214020661SNP117Chr04EdC/T4899932162SNP118Chr04EdC/T5787510563SNP119Chr04FdC/T1084138764SNP120Chr04FdA/G1871957465SNP121Chr04FdG/T2751390066SNP122Chr04FdA/G4814588967SNP123Chr04FdA/G6529469068SNP124Chr04GdT/A1116660169SNP125Chr04GdG/A1744694870SNP127Chr04GdC/A5045446471SNP128Chr04GdC/G6003052072SNP129Chr04GdC/G6800422173SNP131Chr04HdG/A3093162074SNP132Chr04HdA/G6006241075SNP136Chr05AdA/T9003714076SNP138Chr05BdG/T2119437477SNP140Chr05BdC/T8357988778SNP141Chr05CdT/C1769520879SNP143Chr05CdT/C7976321880SNP144Chr05DdC/G1062032981SNP145Chr05DdT/C1870931082SNP146Chr05DdA/C2792921783SNP149Chr05DdG/T7382800384SNP150Chr05EdT/C1705368585SNP151Chr05EdT/C2528432786SNP153Chr05EdT/A5197529887SNP159Chr05FdA/G5781913688SNP160Chr05GdA/G1263685889SNP163Chr05HdG/A1126272490SNP165Chr05HdC/T3511777091SNP167Chr06AdT/C1154390092SNP170Chr06AdC/T7940028893SNP172Chr06BdA/G2696716094SNP173Chr06BdA/G4501990995SNP177Chr06CdG/A1650992696SNP182Chr06DdC/T4767658597SNP183Chr06DdA/G5732582898SNP189Chr06FdG/A1200600099SNP190Chr06FdG/T19208691100SNP191Chr06FdA/C33541175101SNP192Chr06GdG/A8789425102SNP193Chr06GdA/NP194Chr06GdA/G24291447104SNP195Chr06GdC/T31454956105SNP196Chr06GdA/C37677815106SNP198Chr06HdA/G11755209107SNP201Chr07AdA/C30646071108SNP202Chr07AdG/A50071154109SNP208Chr07CdG/NP211Chr07CdG/A38476083111SNP214Chr07DdG/A5344724112SNP216Chr07DdA/G39399976113SNP223Chr07EdT/C51616280114SNP224Chr07EdC/T58995011115SNP226Chr07FdA/G40421759116SNP229Chr07GdC/T30367130117SNP230Chr07GdC/T47823294118SNP231Chr07HdC/A5974408119SNP234Chr07HdC/T42390113120SNP235Chr07HdA/T46002261121SNP237Chr08AdT/C22503753122SNP238Chr08AdC/T30303448123SNP243Chr08BdT/A49018456124SNP244Chr08CdC/NP246Chr08CdA/G57528551126SNP247Chr08DdG/A5783072127SNP248Chr08DdC/A21327050128SNP250Chr08DdT/G40230743129SNP260Chr08FdA/T47714583130SNP262Chr08GdG/A5521298131SNP263Chr08GdT/NP264Chr08GdC/T29360670133SNP265Chr08GdG/A38549239134SNP267Chr08HdT/C29788553135SNP268Chr08HdG/A43204111136SNP269Chr08HdC/T46984854137SNP271Chr09AdC/G52687978138SNP272Chr09AdC/T58344340139SNP274Chr09BdG/A10726372140SNP275Chr09BdT/NP276Chr09BdA/G28210556142SNP278Chr09BdT/A73786852143SNP280Chr09CdG/A35653357144SNP281Chr09CdT/C42194421145SNP282Chr09CdT/C63569281146SNP283Chr09CdC/T70557641147SNP284Chr09DdC/G8593220148SNP285Chr09DdC/T22979077149SNP288Chr09EdT/C16668695150SNP289Chr09EdG/A31441159151SNP292Chr09FdG/C8305974152SNP293Chr09FdC/T21419258153SNP295Chr09FdT/C35203776154SNP296Chr09FdG/A62895631155SNP297Chr09GdA/G29917254156SNP299Chr09GdC/T58110898157SNP300Chr09HdC/A6518299158SNP301Chr09HdC/T12810399159SNP304Chr10BdA/G6500443160SNP307Chr10BdG/A52600442161SNP308Chr10CdT/NP309Chr10CdA/G25935024163SNP310Chr10CdC/T40676298164SNP311Chr10CdC/A52537986165SNP312Chr10CdG/A62369477166SNP313Chr10DdC/T7759159167SNP314Chr10DdC/NP315Chr10DdG/A37706115169SNP318Chr10EdC/T26137885170SNP320Chr10EdG/T50872486171SNP321Chr10EdA/T55099959172SNP322Chr10FdT/C8402636173SNP323Chr10FdC/T41296315174SNP325Chr10GdC/A28266076175SNP326Chr10HdA/C12314516176SNP327Chr10HdC/NP328Chr10HdG/A20326048

（资料性）

多重PCR引物、连接引物多重PCR引物信息见表D.1。多重PCR引物信息引物名称序列长度（bp）PCR长度(bp)100FGCAGGACGCAATTCTGTTATCAT23100100RTAGTGACTTGTTGGCATGTTTGTC24101FGCCACAACATTCAAATACCCACA2391101RAGAGCCGCTTGAGCTAGCAAT21102FGAACTATGGAAATTTAGGTATTTGGTGCT29108102RGAGTATTGATCAATAAAAGACGGTGTGT28103FCCTGGCCATATGTTCTAAGGATAGC2598103RATTGACAGAGCAGTAGTGCCACCAG25104FGTCTGTCCTTAACACTCCTGCTCCTT26181104RAAGTGCACATGTAGTGCAACAACC24107FTTCCATGCGTAACTGTCATAGCTG24107107RTCGTCCATGTGGTGTGACCAGTA2310FGTTTGTTCACACCAAAATTGAAGGA2512410RCAAATGAATGATCAAAGTGTAATGTAGG28110FGGATGATTTCAAACTTTGGACGTGTAG2790110RAGATTTTACTGTTTGGTTGGATGCTTG27117FGATGCCAAAATGATAAAGCACTGCTAC27115117RAGGAACTTTGGGCTTCTTATTCCAG25118FGAAGATAAGTTTCAAGGCACTGGTTTAG28123118RCGAGTACCCTGTGGCAAGAGCTA23119FGAACCAGCAGATCGATATATGTTGAAG27108119RATGCACTTGGACATGCATGGAGTA2411FTAGTCGAAATGGATGGTACAAAATGC2612111RTGATGAGTTGATGGGGCAATACA23120FCTCTAGTGCTTAATTGCTTATTTTTGTACC30130120RGGAATGTTCGATGATCCTTACCAAT25121FTGAAATGGTTGTTTAGTTATCTGGTAAG28112121RAACTACACATTTGGAAGACAGATAAATG28122FCCCATTTCTACTTTCTATTTGCAATGTC2891122RTGTCAACAAAACCATAGGAATGCA24123FTTTCGGTTAGCCAGCGTGATTA2297123RCCCACTTGGCTTGGAGACAC20124FAGGTACTTAAATTGGCCCCTGGATAG26102124RAAGCAGGGATGTGACAATTAATCAAG26125FGTAGCTTGTTGTACCTTACCGCATCTA27130125RAGCTGAAAGATTAACTTGAGGAGCCA26127FCACTTCGTTCAATTCCGAGGCT22108127RATGGCGATGCAGACATTTCAG21128FCAAAACATGTGCCACCGTACACTA24116128RATCACGTAGCTCGTCTCCCCATT23129FATGAGGCGCACATCATATAAAGATT25188129RCGTTAATTAGCATCTTGCCAAAAAAGA27131FTTGTAACTGGTAAAACTTTCCAGGTAG27113131RGCAGAACCAGCTAGAATGAGAATCA25132FATGACAAAGGAGCCAAAGCTTCA2393132RGCAGTACACGTTTAGCAAATCAAGAC26136FCATGTGTCATATAAACAGGGGAGCA2596136RACTATATATAGCTAGGCCGGGCACA25138FGCACAAGTAATCCAAAGTTTGAGGTC26101138RAACCGGCCACTTGATATCCATC22140FATTTTCTATCCACCACATGGAATCTG26149140RAAAGGAGTTGGAGGTTGGGACT22141FGTAAGTTTTCTTATGCTGGATTTGC2578141RGCGTCTGATGTTCTAGTCAGTGTGA25143FTCGACCTGTGCCGAATGCT19136143RGCAAATTATGTACTCAGTTATCCACCA27144FATCATTTGAAATTTTACGAGCGATA25137144RGACTGATTTAATATATTGAGGCATTTGA28145FCGGTGCATAGCACTCATTTCAAC23104145RGGAGGTTGGCAGAAAATAGAATGC24146FGATTCAAACATTCAAGCCAATACCTTAC28147146RTTCAATGTAGCAAAGCCTCTAAATTCAC28149FAAGAACGGGTAGGGCGATTCC21100149RTGAGATATGAGGGGATTAAGCATC2414FAGTTTGCGACAAAGTTAGCGAAAG2413614RGACGACACTAGTTTACTGCCCAAG24150FTCCAAAACATGCTAATGGAGATCG24127150RCTCCTATCAGCATCAGCAAGGAAA24151FGCTTGCGGAAATAGTGAGCTTG22122151RGTCTGATTCTGGTACAGCTCTTAGGT26153FGCTATGTTCTTGGCGATCAACTC23118153RGCATCGACCATTACCTACTACGCT24159FCCCAACCTAATGGAGAACAAGGTT24154159RGCCAAATGGCTCAGCTAGGGT2115FAGGGGGGAAAAGGAGGTAAAGTTG2413615RTCTTCAGAAAAGAAGAGACCCCAAT25160FTAAACATGGCTAGCAGGGTGTGTT2483160RGGGGTAAAATTACCCACTTGAGGA24163FGGTTTGTGAAGGTTGATTTCGAG2394163RTTAAAATGTGTGACAACTGCCACTTTA27165FTTCTTCACACGAACTAGCAAGCATT25145165RCGATACACAACTGGTAGATGGAGAGTC27167FATGGATCCCAATAGATTAGTGCACAG26111167RCTGCCATGAGAGTTCCCAAATTC23170FCTGTCACCTATGTATTTCACATAAACAA28151170RGCTCCAAAGTACATTGATTCCCTTAG26172FCAGGTATCAACACTAAGCTATGGACCT27165172RATATCCTGGCTGAATGCTAGAATGG25173FCACACCTGCAATCAAAATAAAGTGTTAG28137173RCCTACTTAGCTTCTTCTTGACCGATG26177FAGCTCTCGGTGCATTCAGTTCA22124177RGCTGTCCATTGAGCAGGATTGTA23182FGGTAGCGACGAGGAACGTGA2074182RCAGCTGTTGACAAGTGCAAGCTC23183FAGGCCCACGGCTTACCATTAAC2280183RCCCAGTGCTCAGGGGACTAAA21189FAGTGTAGGTTGCAGAGGAGCCAAT24155189RAATCGCAAGGTGCGGTAGTG20190FCTTCTATCCATTCCGTTAGAGTTTGC26101190RATATTAGACGCGCACTTGGTTGA23191FGCTTCTTTTGCACCTGGTTACAGAC25107191RAACTACAAGAACAAATTGAGGCAAGAT27192FACTGTATGACCCACATGGAATAACATT27142192RGAATAGGGATGCTCTATGCTGCTGTA26193FCACATTGTTCGTGCTGTGGATAAC24130193RGACCATCCAAGAATGTCATGAAAAG25194FAACCACATATCACATCGGCATTC23103194RGTGAATGACTCTGTAGGAGCTGCAT25195FACCAGAGCCTGTGACGAATCA2181195RATGTCCGCTACTGGCGTTTC20196FCGGAAAGGTGTGGCGAGATAA21143196RCGGCCAGGAGATGTCAAGGA20198FGTGTCTTGCTTATGTGGCAAGGATAG2694198RCTAAAACTCACTATTTTGCGGGACA2519FGGTGAATCGCCCACAACTG1912919RGTTTGTGCTTGTTACTCTTGGTGATCAT281FTCTAGATGCCTAGGTTAGAGGCCAAT261141RGCATGTTCTTGGGAGCGTCTA21201FCACTCGATTGTATCACCGGTCTG23102201RACGCACTGAGGGATAAGTAACCGTA25202FCCGGGAGAGATGTGAGAGAGAAC2381202RCAGATGTTCGTCGAGGTACAGATTC25208FATTTGCAACCTAAAATGCAAGGAGT25113208RTTTCAATAAGTCAGGTAGCATTGTG25211FCTCTTGTTGAGCTTGAACTGGCAAT25137211RCAGGTGAGCAAGGTCAGGACGTA23214FATGCAGTGCTTGAAGAGAAATGTAGTG27128214RCCCTCCTTTCTCAGATCTTTTGAACA26216FCTTGGCGAACACCATCTCAATTA23132216RCTGGGTTTTAGGGCTGCCTTC21223FGTGTCATCCCCTCTCGAGTCAC22160223RTCCTTTGGACAACAAAAAGAACGA24224FGAGGTTTCTGCTTGGGCTCAG21120224RCTTGCACTCAACCTGCTAGGAAAT24226FGTAACTAAAGCCTATCACATCTGGCTA27131226RATTATAGCCCAACCTACCACCATTCT26229FATACCGCTGCAGCTCCAGAAAC2287229RTCCAGTTTTCCAACAAGTACTGTCATC27230FCTCATAGAGATTGGAGGGGAATGAGAG2773230RAATCCCTCCTAACCCTTCTCAAATG25231FAGCAACAACACTGGATTGCTTCAC24134231RACTCGCTGAACAAGATTGTCGAAAC25234FCCACTCCCTAACCTGACGCTGT22154234RGTGACGCAAGCAACTGAGAGAAC23235FCCGAGGTTGGTAGCTAGCGTTT2287235RTGCCTCTATATAAGTGCAACCAC23237FAATCAAGAGTGTTGGGCAATCCTA24108237RCCTCGGGGAGATGGCTGAAT20238FGGCGTATACTCTGATGCATTTTCAAC26140238RTGCAAGAAGCTAACTGTCAGTCAAAG2623FCTTTATGGTGGGGAAGAGGATCAAG2513823RGTGACTGATATCCCATCCACATTCTC26243FCATCAAGTATTCTTGGGCGGAGATA25141243RCCATTTCTAGACATAAATGTTGACGCTATT30244FTTCACCTTTAGAGTGACGCACTTG24153244RTACTACTGGCCAACGACGTTTCG23246FCAACTACAAAATGGTCTTAGAACTACAC28107246RCATCTGCAAAACATGCACCTGATATAC27247FGGCATGAATGTATTAAGGTCTACAATC27149247RGACATGGTTCGTAACACGGAAAT23248FGTAATCGGATTGGCCCATTG2086248RGGCACTGAACAGAAGATTGCAG22250FCCTATCAAATAAATACTTGGCCCACA2684250RGAGCCCAGATAGGCAAAGATCA2225FAGACAGAGGAAGACCGATGCA2118925RGACGGCAGGGAGGTTGAT18260FAAAAAACTTACCTCTGTGCGCATTTA2691260RCTGCTCTAGCCAATCTCGTACCAC24262FCTACTTCATGATGCTTTGCACGA2389262RACGACGAACAGAACGCTGATGAT23263FTCTGATACGAATCAAATGCACGAT24116263RGATTAGAATTTGTGGTCTGCCTCAG25264FTCATTGACCAGCCAATCTTGC21105264RTGTGCAAGTAAATTACAAGAGCGAAC26265FAAATTAAGCCGAGTATGAACTGATTG26103265RCTAGGGGCAACCGTGTTAAGTG22267FGCCATAGTGATGGTGGCACATAG23122267RCGTCTGAAATCGGATGTCTAGGAAT25268FCCACATACTATTCATAACTAAAGCGTTG28101268RGTTTCAAATTGGTGCGTGGAT21269FCTTAGATTGAAGATTGCTACACATAGATT29120269RGTATTCTTCATCCAAGAGTATATTGAGGT29271FTGTGTAAACCGGTTGGGATCTTC23163271RGGAACGTGGTACTCTACGTCTAGTTTG27272FATGTAGTGGAGGATCAACCCATGAT25157272RTAAACTGCCCTCTGGCTTGCGACA24274FATATCGTAGATATGCCTCTGCTCTGG2686274RCTTTCCTATGCTTCAAATGATACGACT27275FATAGTACTGAAATTCTTTGTCTGTGTAGC29110275RTGTGTAGTGAACAAATTCTCGCATAATC28276FAGGGGCACAAAGAGGTTGCTCTA23143276RTTATGTCCGATTGAGAAATCCAACTTC27278FCGGCGACATCCACTCTTTACA21145278RCATCTGTATCGTTCTCTGATGAATTGG27280FTGCAAATGCACTCCTACGGTACTC24127280RGGCTTGCTGGATGCAACTGT20281FTTTCTGGACCATACCGACCACTG23151281RAAGCTTGATATCACATAGAGGCACAT26282FGATGATTATCCCAAGATTGCCTTG2497282RCACCTCGCGTCATACAACGTCT22283FTTGCTACAATATTTGCGGAGGAAG24124283RGTAATGGACTAGTTTATCAATTCCTACTG29284FCCTGTCCATTCGAACCCAATATG23128284RAGACGGGAAGTGGGCTGTAAATAG24285FAAGTGCGTGGTAATGAGATCCTG23194285RGAAACTCAAGCTGCAACTCAGTCA24288FCTGAAATTCTCCAATAGGCTGTTGAT26167288RACGTTACCACATCAGCAAAATACCAT26289FGGCTCCTTACAAATCATGGCAAC23162289RACCTCAGCACTCTTGACAGCATC2328FTAGTGCCATATGTGTATTGTACCGTTC2712028RCAATCATAACTTACTATCAAGATGAACTG29292FCAACGTTGCACTTGCAGGAGT21149292RACGACGGACGGTGACTCAAG20293FGAGGACAAAGGAAACCCATACGTG24191293RGTCTATCCAACTCGATCTCGTGTCA25295FTTGGTATTAGGAAAAGGCACATAGTTC2770295RTGTGCCTGGGAATATGGTTATGTC24296FGAGGATCCACAAATTGCAGCTCT23115296RACTATCCAAGAGCATACGCACGAC24297FAAGGCATACTGTGTACAACTCCCAAT26160297RGGCTAGAGGATGCTACATCTTTGGTG26299FGGCAATAAATTAAGCTCTGGGTGT24205299RGGTGAGAAGACAAACCTAATCATCA252FGTAGGAGATCAATAACGACGAGACAAA271622RCCATGACGACGCAATAGTTGGTAC24300FCTCGTCCTGGACTTGGAGGTAG22121300RATGCTGATACCCAAATATGAACTCCA26301FATGAACGTTGACGTCGGATGC21125301RGAGAGGTTCTAGAGTTGCTTTTCATCA27304FCCTTCCAAACGTAGAGGCCATACT24128304RAGTTAGGGATCGAAATAGGACATCTG26307FAGTGGTTTTCAAGTTTGGACCTCA24117307RAGGTACCCTGGGTCCATATCAC22308FTTACCCACAACTAGTGCCACAACA24110308RGATATCAATGAGTGTGCTGCCAAAC25309FGTTGCATGTGAAGTCATTGCTTATG25170309RAGCTCTAATAAAAATAGCGGCAAAGC26310FGGCGTTGAATTAGTGATAGGGATTG25139310RCCTCTATCTGATATTTGCCCAACAAC26311FATACTTGCATATACGTGGAACCAGAG26167311RCAAAGAGCCGATCAGCCAGT20312FGGACAGATGCCGGGCTAAGAT21113312RGAACCTGATAACCTAATAACGGTGGA26313FTAGAGAAGGGTTTTTTTCTTGGTAACT27158313RCACATTAGTGACGGTTCAAGATGTA25314FGACCGACGGTGATGAGAAACAG22117314RCCAATCCTGGTGTTCCAGTAGGAG24315FTGGTACTCCAACACACCACTGATAAT26160315RGTCGTTTTGATCAAGTTGGCTCAG24318FCATCGTAGGTAGATGATCTCAACTTTT27165318RTTGGTGTGTTGCCCAATTCAG2131FACGAGTTCCAGGTGATGCAGA2115931RGCAGCTGAACGAGCTGTTCCTA22320FAACTCTGAAAATGACCAGGTGCTAAC26111320RGCTTTGGTAGAGGAATCATTGGTG24321FTTCAGAACTTCAGCGTATATGCACCT26138321RCACCTTTAGTCAAGCTGACCCTGTAAC27322FCTTTATTGGCAGCGACGACCTA2270322RCCCGTGCTAGCTGTCGAGGT20323FGACACGCCGGTCACAGAGATA21105323RCAGGTAAATGCCATCACCAGAGTC24325FTCCATCACTTGATCCCAACGAG2283325RCCTTCTAAGCAGACACAGATTCTACTA27326FCAAGAGAGTTACAAAACCGACTTTACA27144326RGTTGTTGTTGCTGTAGGGGCTAA23327FCTGTGTGCTGGTTTTATCCGTTTATAG27144327RACACGCTGTGCATCAATCAAGTG23328FATCGCCATCCATCAGACACG20120328RCGTGCGACAGACGTGTCCTA2032FCTTGTGCTGATTGCTGCTGCTA2214732RCTTTGTTTAATTCACACGATATCATT2634FTGTGGCGGTGTGGGTAGCA198934RGCTGCTTGGTGCGGAGCTT1936FCTTTGACTTGGGCATCGAACT2110036RTCGCTCCCTCCACACTACG1937FGCCAAATTCCGGTATACGCA2013937RTTTTGTCCACGTTATATTTTCCTCG2539FCAAAAATTGACACACCTTATTTACAGG2714139RGAAAAATTATCAGTTCATGAGGGGAC2640FCCTCTGTGTGTTTCTTCCAAAATACC2616640RCCATGTACCAAATTTACATCGTTTCA2642FTTGGGGCGGTCGATTCTCG1911142RAACGGAGCAAGCTATCACTCACC2343FGACAAGGGACATAGTTGAGCCAAA2414343RGTGCTCTTATAATTTGGACAGTTCTGTTAC3045FCAGAAACCACACTGTTCTAGACACGA2610845RCGAGTTCTCTGGAGTTGCTGG2146FCACTACAAATATCATTCTGCTGAAGAC2713046RGTCAGGAGATTTGAAAGCAATATAGATG2848FCGAAACACTTGGCTCTATTATTGAACTTAC3012648RGAACAACATGCAGCATCCAAAGTA2449FAAAACGACATTTAAAAAGGGACCAG2512749RTTATTAAGTTATTACTATTACCGTGGCTA294FGGAACCTTTTCGGATTGTCAG212984RTGCAGGTAACGTGATGAGTAAAACTG2650FTGGTTTATGCATTAGTTACATATTTGG2710350RGAGTTCAACAGTTAATAAACAACATACAGA3052FGGCACAAGGAAGGCAAGAGTG219152RTTCACCTGTTCGCCTTCCTGTA2253FCATGCTGGAACACACAGGAAGATA2415953RTAGGACATCCCCTAACCAGTAGTCTAT2755FTTTTAGAAATCACATCTTATATTATGGAAC3013155RTCTGGAGCAAGTTTGTAGAATTGTACTT2859FCCCATTCTGCTACACTCTTCTCCATAC2714959RGATTCCTCAGTTGGCTATAGTTGGC2560FCTGTGAAGTCCCTGTGTTCCAAAC2415860RATGCCCAAAGCGGCTATTAC2062FCAACGTCTTGTTGGTGAACGA2117962RCTCAAGATAAGGAATGAATGTCAAAGA2763FGGGAAAAAAACTCAGATCTAGCTTACC2711463RAACTTAGAGAAATTTGACTCTGGATGC2765FACCCAGCGATTTAAGCAACAATTC248965RCAACTCACACTGAAGCAAGTGTTTT2566FTGGGGGAAAGAACAAGAAAGTTTG2411366RTCCTAACAGTAAGTTGTTTCTCATCC266FCTGGCGCAGGTAAAACCCTA20886RGTAGCGCCGTCGTTCAAGA1970FATGTCCCATTGTAGAGCGCTGAT2317370RAGCAAATGCAGATGATTTGAAGAA2471FGATGACATGTAGATTAGTTGAAGAGATT2813671RATTCACATGGTTGCTATGCTACTC2476FGATGAAATTGCCTTAGCCTCATATTTC2712976RGTCTTTGCTATTTGGTTAGTACTAGGTC2877FCTTAGCTTGCGCTACCGTGT2012477RCCATCTCTTAACTTTTATTATTGATTTCC2979FGAACAAGTTATTTGATTCTGACCCAT2611479RGATAAGGTGCTTACTCTTGCTGGT2482FTAATCCGACAATGGATGACACC227282RTGGCGCTGCTCCTTGTAAC1984FCGGTGTCTATCCTAGCGCTTCA2214284RAGGCATTTGATGTACATGTGCTG2385FTGAGTCTCCCGTGAGCAGCAG219985RCGCCATCGGGACCATTTGA1986FGATGATTTGGTTGCACACTTGA228586RGTGAAAATAATTCGCTACAAACCTTC2687FCCAACCACGTCATCGACCTC208087RGTCCTGGCAGGGGATATTGC2088FCCCTAGCCGTTGATTTGGTG2012988RCATGTGTCCTAAGATAGTTGGATGG2589FATGAGCGAACCAAGTCGACAAT2210089RGCAATAGAACCTGCGAGCTGTG228FCTAACAAGGGTAGGAATCGAGAGAAA261488RAACAGAATTCAGAAAAAGGTTGTCAC2690FTGTACCAGGCATTTGCATCTCTG239090RTCGCAGATCCAAGCCTGAAAATAC2491FGGTTATGGTCTGAAGCGGTCATT2312791RGTGTTCCATGATTATGAATTTTTTAGC2792FGATATTTTGGCTTTGTTAGATATACTG2715192RCACAAATCCTTATATTGTACTCCCTCAAT2995FGCACTCCTTTCGCTGAGCATAC2217895RGGGGACCCTTCACTATTCATTCA2396FTGCTGCACCATCAAAATCAGC219896RTGCATGCCTGCGGCTACTG19连接引物信息见表D.2。连接引物信息引物名称序列(5'--3')长度（bp）100F3AAAGGCAAGAGTCGACAAACATGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[NED]53100FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTAAGCTTAAAAGTAATTTGGTGACATCTTTGAC73100FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTAAGCTTAAAAGTAATTTGGTGACATCTTTGAG75101F3GCAGTTGCAGCTTAAGCATCTTCTCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[FAM]59101FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTATTCAAATACCCACACTAACCTCTAAATGAC72101FTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTATTCAAATACCCACACTAACCTCTAAATGAT74102R3AAGATCGAAAGCACCAAATACCTAAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[VIC]59102RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTACTAAATGAGCAACAATAAATGCTCAAATGACTGG77102RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTACTAATATGAGCAACAATAAATGCTCAAATGACTGA79103F3ATCCTGGTGGCACTACTGCTCTTTTTTTTTTTTTTTTTT[NED]39103FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTGATAAATTAGCTGGATGCCTGGAGAA67103FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTGATAAATTAGCTGGATGCCTGGAGAC69104F3ATGATTGGGCATTTTTAATACGTGTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[NED]59104FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTTATCGTGTTTTGGAAACTACCTTTAATATGCGTA77104FTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTAATTATCGTGTTTTGGAAACTACCTTTAATATGCGTT79107R3AAATGCTGTAGTCCTGTAGCTATCAGTAACTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]56107RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTGGGAACATGCACGATGCCATG62107RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTGGGAACATGCACGATGCCATC6410R3ATAAAATGGACCGAAGGGAGTACTTTACTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[VIC]5710RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTCGTGTCATTCTAACTTGCGCAAG6410RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCGTGTCATTCTAACTTGCGCGAA66110F3ATAAACATTGCCAAGCATCCAACTTTTTTTTTTTTTTTTTTTTTT[VIC]45110FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTGGTCTAGGATCAAGGTCTGCAGCGTA68110FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTGTCTAGGATCAAGGTCTGCAGCATG66117R3TCTCCATGGCTTCCTGTAGGTAGTTTTTTTTTTTTTTTTTTTTTTTTTTTT[VIC]51117RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTGAGGAGTATCTGTATTTGAAGGGTGTACG70117RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTGAGGAGTATCTGTATTTGAAGGGTGTGCA72118R3CAAACTGAACGCCTAAACCAGTGTTTTTTTTTTTTTTTTT[NED]40118RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTCTCCTATCATCCTCAAACCGATTGG66118RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCTCCTATCATCCTCAAACCGATCGA68119F3CATTTCCATGGCTACTCCATGCTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]48119FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTACAGTGGATCGGATCATATTCCAGAAGTC70119FTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTACAGTGGATCGGATCATATTCCAGAAGTT7211R3AGCTGAAAGGATGCATTTTGTACCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[NED]5811RATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTAATTCAATTCAGTGATAAAATAAATCTCTGTCT7511RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTATTCAATTCAGTGATAAAATAAATCTCTGTCC73120R3AAATAAAAGGTACAAAAATAAGCAATTAAGCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]64120RATGTACGTGATACTGACAAGTTGTACATGATATACACATATTACTAAAAATTCGCTTTAAACTATATTAATACTCTTGTT79120RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTACTAAAAATTCGCTTTAAACTATATTAATACTCTTGTC81121R3AAAGTTATGTCCTTGTAATTGTGATATTAATGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]70121RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTAAATGGGATTTTCAACCTGAAAAGAC68121RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTTAAATGGGATTTTCAACCTGAAAAGAA70122F3CTGTTGTTCAATACACTTTGCATTCCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[FAM]58122FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTAATGTCGTTCTTCCACACAATGATGAA68122FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTAATGTCGTTCTTCCACACAATGATGAG70123F3TCTCGACCCTACACACGTGTCTCTTTTTTTTTTTTTTTTT[NED]40123FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTATTCACCTTCCCTCTAGTCCGACA66123FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTTATTCACCTTCCCTCTAGTCCGACG68124R3CAGTGGTCACTATCCAGGGGCCAATTTTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]53124RATGTACGTGATACTGACAAGTTGTACATGATATACACATATTCCGTTCTAGATCAAACCCTACTGATCAGT70124RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCCGTTCTAGATCAAACCCTACTGATCAGA72125R3GCTTGAAAGTGAGCACACATGCATTTTTTTTTTTTTTTTTTT[PET]42125RATGTACGTGATACTGACAAGTTGTACATGATATACACATATTCATCAGGTGAGGTTCTTTAGGGAAT66125RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCATCAGGTGAGGTTCTTTAGGGAAC68127F3CAGCGATGAAGGTGAATGCCTTTTTTTT[NED]28127FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTAGCGCAATCGGATCAGGAGCAA63127FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTAGCGCAATCGGATCAGGAGTAC65128F3TAGCGCAATGGTAATGGGCTTTTTTTTT[VIC]28128FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTCACTAATCCCCATGCTCCCTGC63128FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCACTAATCCCCATGCTCCCTGG65129F3TGCACGCACACTCCAATTAAGTTTTTTTT[NED]29129FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTACGGCCGTGCATTAGACCACC62129FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTACGGCCGTGCATTAGACCACG64131F3TTCGTCGCCGTATCCCCCTGATTTTTTTTTTTTTTTT[NED]37131FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTGTTTTGCAGCTACTCCGGACA64131FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTTTGTTTTGCAGCTACTCCGGACG66132F3GTTTTTTGTTTTTTTCTTTTGGTCTTGATTTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]69132FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTCTTCAGGTAAAATTATGTCCCAGTGCAA69132FGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCTTCAGGTAAAATTATGTCCCAGTGTAG71136F3TCACATGTGCCCGGCCTAGCTATATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]58136FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCTGTAATTTCCACCAAGATTCTCTATCATGCA75136FTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTATCTGTAATTTCCACCAAGATTCTCTATCATGCT77138R3TTTTGATGGCACGGACCTCAATTTTTTTTTTT[NED]32138RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTCTGCCTAAGCTGAACTTCCCAAC64138RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCTGCCTAAGCTGAACTTCCCGAA66140F3TGTGCATAACATATGTTTGCCCATTTTTTTTTTTTTTTTTTTTTTTTT[PET]48140FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTCTGAAAAATGCCATGCCACTATGC65140FTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTCTGAAAAATGCCATGCCACTACGT67141R3TATACTAGAATAAGTTGCAAATCCAGCATAATTTTTTTTTTTTTTTTTTTTTTTTTTTT[FAM]59141RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTATGTACTAATTCTAGTCAGTGTGAACTTTGCCAACG77141RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTATGTACTAATTTCTAGTCAGTGTGAACTTTGCCGACA79143R3TAGCATTTGGTTGTGGTGGCTTTTTTTTTTTTTTTT[NED]36143RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTGCTATCATCTAGAAGCCGGCTACG65143RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTGCTATCATCTAGAAGCCGGCTGCA67144R3CTATTGGATTATCGCTCGTAAAATTTCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[PET]59144RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTAATACGTTGAAGTTTGTAGAGTCGAGCATCTAG74144RGTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTAATATCGTTGAAGTTTGTAGAGTCGAGCATCTAC76145R3TTCTAAAATGCAAATCATAATAACTATGGTTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[FAM]67145RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTATAGAATGCTTGTAAGTACATGATATACATTAAAAGAG79145RTTGTACGTGATACTGACAAGTTGTACATGATATACACATATTTTATAGAATGCTTGTAAGTACATGATATACATTAAAAGAA81146R3TGTAATATGTTACAACAATAAATGCATAAATATAGTATGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT[VIC]75146RATGTACGTGATACTGACAAGTTGTACATGATATACACATATTATGTAGCAAAGCCTCTAAATTCACAAAAACCT73146RCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTGTAGCAAAGCCTCTAAATTCACAAAAATCG71149F3CTCCTGTATACCAGATATTCCTGTGCCTTTTTTTTTTTTTTTTTTTTTT[VIC]49149FATGTACGTGATACTGACAAGTTGTACATGATATACACATATTTCCAATGGGAATCATGATCGTGT64149FCTGTACGTGATACTGACAAGTTGTACATGATATACACATATTCAATGGGAATCATGATCGTGG6