Q/LB.□XXXXX-XXXX前言本文件按照GB/T1.1—2020《標(biāo)準(zhǔn)化工作導(dǎo)則第1部分：標(biāo)準(zhǔn)化文件的結(jié)構(gòu)和起草規(guī)則》的規(guī)定起草。請(qǐng)注意本文件的某些內(nèi)容可能涉及專(zhuān)利。本文件的發(fā)布機(jī)構(gòu)不承擔(dān)識(shí)別專(zhuān)利的責(zé)任。本文件由福建農(nóng)林大學(xué)、廣西大學(xué)提出并宣貫。本文件由廣西標(biāo)準(zhǔn)化協(xié)會(huì)歸口。本文件起草單位：福建農(nóng)林大學(xué)、廣西大學(xué)、廣東省科學(xué)院南繁種業(yè)研究所。本文件主要起草人：趙新旺、張木清、鄧祖湖、暴怡雪、趙薇彤、黃江鋒、王勤南、姚偉、徐良年、吳嘉云、胡琴、張南南、張垂明、余凡、蔣洪濤、肖勝華、張偉。FORMTEXT甘蔗品種SNP分子檢測(cè)技術(shù)規(guī)程范圍本文件規(guī)定了構(gòu)建甘蔗(Saccharumspp.hybrid)品種SNP分子標(biāo)記指紋圖譜的原理、檢測(cè)方案、檢測(cè)程序和結(jié)果判定。本文件適用于構(gòu)建甘蔗常規(guī)品種的SNP分子標(biāo)記指紋圖譜和身份鑒定。規(guī)范性引用文件下列文件中的內(nèi)容通過(guò)文中的規(guī)范性引用而構(gòu)成本文件必不可少的條款。其中，注日期的引用文件，僅該日期對(duì)應(yīng)的版本適用于本文件；不注日期的引用文件，其最新版本（包括所有的修改單）適用于本文件。GB/T3543.2農(nóng)作物種子檢驗(yàn)規(guī)程扦樣GB/T3543.5農(nóng)作物種子檢驗(yàn)規(guī)程真實(shí)性和品種純度鑒定GB/T6682分析實(shí)驗(yàn)室用水規(guī)格和試驗(yàn)方法GB/T19495.3轉(zhuǎn)基因產(chǎn)品檢測(cè)核酸提取純化方法術(shù)語(yǔ)和定義下列術(shù)語(yǔ)和定義適用于本文件。品種真實(shí)性驗(yàn)證varietyverification與其對(duì)應(yīng)品種名稱(chēng)的標(biāo)準(zhǔn)樣品比較，檢測(cè)并證實(shí)供檢樣品品種名稱(chēng)與標(biāo)注是否與標(biāo)準(zhǔn)品種樣品相符。多重PCRmultiplexPolymeraseChainReaction，mPCR在同一PCR反應(yīng)體系里加上二對(duì)以上引物，同時(shí)擴(kuò)增出多個(gè)核酸片段的PCR反應(yīng)，其反應(yīng)原理，反應(yīng)試劑和操作過(guò)程與一般PCR相同。

連接酶檢測(cè)反應(yīng)LigaseDetectionReaction，LDR在TaqDNA連接酶的作用下，當(dāng)左右兩條寡核苷酸探針與目的DNA序列完全互補(bǔ)，并且兩條探針之間沒(méi)有空隙時(shí)才能發(fā)生連接反應(yīng)，最后通過(guò)熒光掃描片段長(zhǎng)度，實(shí)現(xiàn)對(duì)SNP位點(diǎn)的檢測(cè)?？s略語(yǔ)下列縮略語(yǔ)適用于本文件。bp：basepair堿基對(duì)CTAB：cetyltrimethylammoniumbromide十六烷基三甲基溴化銨DNA：deoxyribonucleicacid脫氧核糖核酸PCR：polymerasechainreaction聚合酶鏈?zhǔn)椒磻?yīng)LDR：ligasedetectionreaction連接酶檢測(cè)反應(yīng)SNP：singlenucleotidepolymorphisms單核苷酸多態(tài)性?xún)x器設(shè)備及試劑儀器設(shè)備及試劑見(jiàn)附錄A，所用試劑均為分析純。試劑配制所用水應(yīng)符合GB/T6682規(guī)定的一級(jí)水的要求。溶液配制溶液配制見(jiàn)附錄B。SNP標(biāo)記一套甘蔗品種真實(shí)性檢測(cè)SNP標(biāo)記包括177個(gè)核心SNP標(biāo)記、位點(diǎn)及物理位置見(jiàn)附錄C。引物信息引物分為多重PCR引物、連接引物見(jiàn)附錄D。檢測(cè)程序樣品準(zhǔn)備采集甘蔗葉片樣品重量應(yīng)不低于3g，并用自封袋包裝，冰盒攜帶，4℃條件下保存。用混合樣品檢測(cè)的，每個(gè)個(gè)體上等量取樣，混合后提取DNA。采集種莖的，常溫運(yùn)送，溫室種植，待長(zhǎng)出真葉后采集葉片取樣，方法同上。采樣脫毒組培瓶苗的，在提取DNA前不開(kāi)瓶，防止污染。樣品在4℃下可以保存2周，在-20℃下可以保存6個(gè)月，在-80℃下可以長(zhǎng)期保存。設(shè)標(biāo)準(zhǔn)樣品，與樣品采集同樣方法。從供檢樣品中分別取出有代表性、符合GB/T3543.2規(guī)定的試樣，采用混合樣或單個(gè)個(gè)體進(jìn)行檢測(cè)試樣來(lái)源應(yīng)至少含有3個(gè)個(gè)體。DNA樣品準(zhǔn)備參照GB/T19495.3的方法或者使用試劑盒進(jìn)行DNA提取。保證提取的DNA數(shù)量和質(zhì)量符合PCR擴(kuò)增的要求，DNA無(wú)降解，溶液的紫外光吸光度OD260與OD280的比值介于1.8~2.0，經(jīng)過(guò)瓊脂糖電泳檢測(cè)具有清晰明亮的單一主帶。多重PCR反應(yīng)將配制好的PCR反應(yīng)預(yù)混合液充分震蕩離心混勻后分裝到96孔板里，每個(gè)反應(yīng)孔分裝19μL（表1）。然后每個(gè)反應(yīng)孔里加入1μL的DNA樣本，蓋上蓋膜，輕微震蕩混勻后3,000rpm離心30秒，然后立即將離心后96孔板放置PCR儀上，去除蓋膜，然后蓋上96孔橡膠蓋墊，蓋好PCR儀熱蓋，按表2程序運(yùn)行。177個(gè)甘蔗核心SNP標(biāo)記分為4個(gè)多重LDR體系進(jìn)行檢測(cè)，見(jiàn)附錄E。多重PCR反應(yīng)預(yù)混合液配制試劑表試劑1×100×10×PerfectStart?Taqbuffer2μL200μLdNTP（2.5mM）2.4μL240μLMgCl2（25mM）0.8μL80μLProbeMix2.0μL200μLHotStarTaq（2.5U/μL）0.4μL40μLddH2O11.4μL1140μL總體積19μL1900μL多重PCR反應(yīng)程序變性退火延伸保持循環(huán)數(shù)1step95oC2min12step94oC20sec65oC40sec(-0.5oC/cycle)72oC1.5min-113step94oC20sec59oC30sec72oC1.5min-244step--72oC2.0min4oC1連接反應(yīng)將配制好的PCR反應(yīng)預(yù)混合液充分震蕩離心混勻后分裝到96孔板里，每個(gè)反應(yīng)孔分裝6μL（表3）。每個(gè)反應(yīng)孔里加入4μL的多重PCR純化產(chǎn)物，蓋上蓋膜，輕微震蕩混勻后3,000rpm離心30秒，然后立即將離心后96孔板放置PCR儀上，去除蓋膜，然后蓋上96孔橡膠蓋墊，蓋好PCR儀熱蓋，按表4程序運(yùn)行。連接反應(yīng)預(yù)混合液配制試劑表試劑1×100×10×ligasebuffer1.00μL100μLLigasePrimerMix(1μM)0.4μL40μLDNAligase(50U/μL)1.0μL100μLddH2O3.6μL360μL總體積6μL600μL連接反應(yīng)程序變性連接保持循環(huán)數(shù)1step94oC1.0min56oC4.0min-382step--4oC-上機(jī)檢測(cè)連接產(chǎn)物加入到70μLddH2O，充分震蕩離心后，取3μL與0.5μLLiz500SizeStandard，8.5μLHi-Di混勻，95oC變性5分鐘后用ABI3730XL測(cè)序儀分析檢測(cè)。數(shù)據(jù)分析使用軟件GeneMapper4.1（AppliedBiosystems,USA）進(jìn)行數(shù)據(jù)分析?；蛐团袛嗝總€(gè)位點(diǎn)的兩個(gè)等位基因size大小不同，size大小在設(shè)計(jì)體系探針時(shí)已設(shè)計(jì)好，所以等位基因在峰圖中的位置是固定的，根據(jù)峰型圖即可判斷出該位點(diǎn)的基因型。數(shù)據(jù)記錄方式為具體基因型，數(shù)據(jù)記錄為X/Y，其中X，Y為同一位點(diǎn)上兩個(gè)不同的等位變異，即純合基因型記錄為XX或YY，雜合基因型記錄為XY，缺失位點(diǎn)記錄為NN。單位點(diǎn)基因型判斷的峰圖見(jiàn)附錄F。真實(shí)性驗(yàn)證根據(jù)供檢樣品與對(duì)照樣品在177個(gè)核心標(biāo)記中的位點(diǎn)差異，進(jìn)行品種真實(shí)性驗(yàn)證。將檢測(cè)得到的基因型根據(jù)標(biāo)記順序進(jìn)行排列，并計(jì)算基因型序列的遺傳距離或遺傳相似性。遺傳距離小于0.02或遺傳相似性大于98%的認(rèn)定為同一品種，遺傳距離大于等于0.02或遺傳相似性小于等于98%的認(rèn)定為不同品種?！?dāng)差異位點(diǎn)數(shù)大于等于5個(gè)時(shí)，認(rèn)定為不同品種；——當(dāng)差異位點(diǎn)數(shù)大于1且小于等于4時(shí)，認(rèn)定為相似品種；——當(dāng)差異位點(diǎn)數(shù)小于等于1時(shí)，認(rèn)定為同一品種。

（資料性）

主要儀器設(shè)備與試劑主要儀器設(shè)備PCR儀、全自動(dòng)遺傳分析儀、微量移液槍?zhuān)?.5μL、10μL、100μL、1000μL）、電泳槽及配套的制膠配件、恒壓電泳儀、凝膠成像系統(tǒng)、恒溫水浴鍋、制冰機(jī)、電子天平、酸度計(jì)、高壓滅菌鍋、高速臺(tái)式離心機(jī)、冰箱、超凈工作臺(tái)、紫外分光光度計(jì)、酸濃度測(cè)定儀。主要試劑十六烷基三乙基溴化銨（CTAB）、三氯甲烷、異丙醇、異戊醇、乙二胺四乙酸二鈉、三羥甲基氨基甲烷、鹽酸、氫氧化鈉、氯化鈉、β-巰基乙醇（β-Mercaptoethanol）、dNTP混合液、Ex-TaqDNA聚合酶、10×Ex-TaqBuffer:含Mg2-25mMol/L、瓊脂糖、DNA分子量標(biāo)準(zhǔn)、無(wú)水乙醇、SNP引物及連接引物、去離子水、DNA分析儀專(zhuān)用甲酰胺、DNA分析儀專(zhuān)用分子量?jī)?nèi)標(biāo)Liz標(biāo)記、DNA分析儀專(zhuān)用電泳緩沖液。甘蔗黑穗病菌冬孢子活性檢測(cè)形態(tài)

（資料性）

溶液配制0.5mol/LEDTA溶液稱(chēng)取186.1gNa2EDTA·2H2O溶于800mL水中，加固體NaOH調(diào)pH至8.0，加水定容至1,000mL，121℃高壓滅菌20min。1mol/LTris-HCI溶液稱(chēng)取60.55gTris堿溶于適量水中，加HCI調(diào)制調(diào)pH至8.0，加水定容至500mL，121℃高壓滅菌20min。5mol/LNaCl溶液稱(chēng)取146.1g固體NaCl，加水定容至500mL，121℃高壓滅菌20min。CTAB提取液稱(chēng)取280mL的5mol/LNaCl、20mLβ-巰基乙醇（C2H6OS)、20gCTAB、100mL1mol/LTris-HCl和40mL0.5mol/LEDTA，混合加水定容至1000mL，4℃貯存。SDS提取液稱(chēng)取100mL1mol/LTris-HCl、25mL0.5mol/LEDT、25mL5mol/LNaCl和2.5gSDS，混合加水定容至500mL。5mol/LKAC稱(chēng)取49.67gKAC，加水溶解，冰乙酸調(diào)整pH至5.2，定容至100mL，121℃高壓滅菌20min。1×TAE溶液稱(chēng)取5mL1mol/LTris-HCl和1mL0.5mol/LEDTA，加HCl調(diào)pH至8.0，加水定容至500mL，121℃高壓滅菌20min。75%乙醇稱(chēng)取75mL無(wú)水乙醇和25mL蒸餾水混勻。

（規(guī)范性）

甘蔗黑穗病抗性標(biāo)準(zhǔn)對(duì)照品種核心SNP標(biāo)記位點(diǎn)及物理位置見(jiàn)表C.1。核心SNP標(biāo)記位點(diǎn)及物理位置標(biāo)記物序號(hào)SNP序號(hào)染色體等位基因物理位置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

（資料性）

多重PCR引物、連接引物多重PCR引物信息見(jiàn)表D.1。多重PCR引物信息引物名稱(chēng)序列長(zhǎng)度（bp）PCR長(zhǎng)度(bp)100FGCAGGACGCAATTCTGTTATCAT23100100RTAGTGACTTGTTGGCATGTTTGTC24101FGCCACAACATTCAAATACCCACA2391101RAGAGCCGCTTGAGCTAGCAAT21102FGAACTATGGAAATTTAGGTATTTGGTGCT29108102RGAGTATTGATCAATAAAAGACGGTGTGT28103FCCTGGCCATATGTTCTAAGGATAGC2598103RATTGACAGAGCAGTAGTGCCACCAG25104FGTCTGTCCTTAACACTCCTGCTCCTT26181104RAAGTGCACATGTAGTGCAACAACC24107FTTCCATGCGTAACTGTCATAGCTG24107107RTCGTCCATGTGGTGTGACCAGTA2310FGTTTGTTCACACCAAAATTGAAGGA2512410RCAAATGAATGATCAAAGTGTAATGTAGG28110FGGATGATTTCAAACTTTGGACGTGTAG2790110RAGATTTTACTGTTTGGTTGGATGCTTG27117FGATGCCAAAATGATAAAGCACTGCTAC27115117RAGGAACTTTGGGCTTCTTATTCCAG25118FGAAGATAAGTTTCAAGGCACTGGTTTAG28123118RCGAGTACCCTGTGGCAAGAGCTA23119FGAACCAGCAGATCGATATATGTTGAAG27108119RATGCACTTGGACATGCATGGAGTA2411FTAGTCGAAATGGATGGTACAAAATGC2612111RTGATGAGTTGATGGGGCAATACA23120FCTCTAGTGCTTAATTGCTTATTTTTGTACC30130120RGGAATGTTCGATGATCCTTACCAAT25121FTGAAATGGTTGTTTAGTTATCTGGTAAG28112121RAACTACACATTTGGAAGACAGATAAATG28122FCCCATTTCTACTTTCTATTTGCAATGTC2891122RTGTCAACAAAACCATAGGAATGCA24123FTTTCGGTTAGCCAGCGTGATTA2297123RCCCACTTGGCTTGGAGACAC20124FAGGTACTTAAATTGGCCCCTGGATAG26102124RAAGCAGGGATGTGACAATTAATCAAG26125FGTAGCTTGTTGTACCTTACCGCATCTA27130125RAGCTGAAAGATTAACTTGAGGAGCCA26127FCACTTCGTTCAATTCCGAGGCT22108127RATGGCGATGCAGACATTTCAG21128FCAAAACATGTGCCACCGTACACTA24116128RATCACGTAGCTCGTCTCCCCATT23129FATGAGGCGCACATCATATAAAGATT25188129RCGTTAATTAGCATCTTGCCAAAAAAGA27131FTTGTAACTGGTAAAACTTTCCAGGTAG27113131RGCAGAACCAGCTAGAATGAGAATCA25132FATGACAAAGGAGCCAAAGCTTCA2393132RGCAGTACACGTTTAGCAAATCAAGAC26136FCATGTGTCATATAAACAGGGGAGCA2596136RACTATATATAGCTAGGCCGGGCACA25138FGCACAAGTAATCCAAAGTTTGAGGTC26101138RAACCGGCCACTTGATATCCATC22140FATTTTCTATCCACCACATGGAATCTG26149140RAAAGGAGTTGGAGGTTGGGACT22141FGTAAGTTTTCTTATGCTGGATTTGC2578141RGCGTCTGATGTTCTAGTCAGTGTGA25143FTCGACCTGTGCCGAATGCT19136143RGCAAATTATGTACTCAGTTATCCACCA27144FATCATTTGAAATTTTACGAGCGATA25137144RGACTGATTTAATATATTGAGGCATTTGA28145FCGGTGCATAGCACTCATTTCAAC23104145RGGAGGTTGGCAGAAAATAGAATGC24146FGATTCAAACATTCAAGCCAATACCTTAC28147146RTTCAATGTAGCAAAGCCTCTAAATTCAC28149FAAGAACGGGTAGGGCGATTCC21100149RTGAGATATGAGGGGATTAAGCATC2414FAGTTTGCGACAAAGTTAGCGAAAG2413614RGACGACACTAGTTTACTGCCCAAG24150FTCCAAAACATGCTAATGGAGATCG24127150RCTCCTATCAGCATCAGCAAGGAAA24151FGCTTGCGGAAATAGTGAGCTTG22122151RGTCTGATTCTGGTACAGCTCTTAGGT26153FGCTATGTTCTTGGCGATCAACTC23118153RGCATCGACCATTACCTACTACGCT24159FCCCAACCTAATGGAGAACAAGGTT24154159RGCCAAATGGCTCAGCTAGGGT2115FAGGGGGGAAAAGGAGGTAAAGTTG2413615RTCTTCAGAAAAGAAGAGACCCCAAT25160FTAAACATGGCTAGCAGGGTGTGTT2483160RGGGGTAAAATTACCCACTTGAGGA24163FGGTTTGTGAAGGTTGATTTCGAG2394163RTTAAAATGTGTGACAACTGCCACTTTA27165FTTCTTCACACGAACTAGCAAGCATT25145165RCGATACACAACTGGTAGATGGAGAGTC27167FATGGATCCCAATAGATTAGTGCACAG26111167RCTGCCATGAGAGTTCCCAAATTC23170FCTGTCACCTATGTATTTCACATAAACAA28151170RGCTCCAAAGTACATTGATTCCCTTAG26172FCAGGTATCAACACTAAGCTATGGACCT27165172RATATCCTGGCTGAATGCTAGAATGG25173FCACACCTGCAATCAAAATAAAGTGTTAG28137173RCCTACTTAGCTTCTTCTTGACCGATG26177FAGCTCTCGGTGCATTCAGTTCA22124177RGCTGTCCATTGAGCAGGATTGTA23182FGGTAGCGACGAGGAACGTGA2074182RCAGCTGTTGACAAGTGCAAGCTC23183FAGGCCCACGGCTTACCATTAAC2280183RCCCAGTGCTCAGGGGACTAAA21189FAGTGTAGGTTGCAGAGGAGCCAAT24155189RAATCGCAAGGTGCGGTAGTG20190FCTTCTATCCATTCCGTTAGAGTTTGC26101190RATATTAGACGCGCACTTGGTTGA23191FGCTTCTTTTGCACCTGGTTACAGAC25107191RAACTACAAGAACAAATTGAGGCAAGAT27192FACTGTATGACCCACATGGAATAACATT27142192RGAATAGGGATGCTCTATGCTGCTGTA26193FCACATTGTTCGTGCTGTGGATAAC24130193RGACCATCCAAGAATGTCATGAAAAG25194FAACCACATATCACATCGGCATTC23103194RGTGAATGACTCTGTAGGAGCTGCAT25195FACCAGAGCCTGTGACGAATCA2181195RATGTCCGCTACTGGCGTTTC20196FCGGAAAGGTGTGGCGAGATAA21143196RCGGCCAGGAGATGTCAAGGA20198FGTGTCTTGCTTATGTGGCAAGGATAG2694198RCTAAAACTCACTATTTTGCGGGACA2519FGGTGAATCGCCCACAACTG1912919RGTTTGTGCTTGTTACTCTTGGTGATCAT281FTCTAGATGCCTAGGTTAGAGGCCAAT261141RGCATGTTCTTGGGAGCGTCTA21201FCACTCGATTGTATCACCGGTCTG23102201RACGCACTGAGGGATAAGTAACCGTA25202FCCGGGAGAGATGTGAGAGAGAAC2381202RCAGATGTTCGTCGAGGTACAGATTC25208FATTTGCAACCTAAAATGCAAGGAGT25113208RTTTCAATAAGTCAGGTAGCATTGTG25211FCTCTTGTTGAGCTTGAACTGGCAAT25137211RCAGGTGAGCAAGGTCAGGACGTA23214FATGCAGTGCTTGAAGAGAAATGTAGTG27128214RCCCTCCTTTCTCAGATCTTTTGAACA26216FCTTGGCGAACACCATCTCAATTA23132216RCTGGGTTTTAGGGCTGCCTTC21223FGTGTCATCCCCTCTCGAGTCAC22160223RTCCTTTGGACAACAAAAAGAACGA24224FGAGGTTTCTGCTTGGGCTCAG21120224RCTTGCACTCAACCTGCTAGGAAAT24226FGTAACTAAAGCCTATCACATCTGGCTA27131226RATTATAGCCCAACCTACCACCATTCT26229FATACCGCTGCAGCTCCAGAAAC2287229RTCCAGTTTTCCAACAAGTACTGTCATC27230FCTCATAGAGATTGGAGGGGAATGAGAG2773230RAATCCCTCCTAACCCTTCTCAAATG25231FAGCAACAACACTGGATTGCTTCAC24134231RACTCGCTGAACAAGATTGTCGAAAC25234FCCACTCCCTAACCTGACGCTGT22154234RGTGACGCAAGCAACTGAGAGAAC23235FCCGAGGTTGGTAGCTAGCGTTT2287235RTGCCTCTATATAAGTGCAACCAC23237FAATCAAGAGTGTTGGGCAATCCTA24108237RCCTCGGGGAGATGGCTGAAT20238FGGCGTATACTCTGATGCATTTTCAAC26140238RTGCAAGAAGCTAACTGTCAGTCAAAG2623FCTTTATGGTGGGGAAGAGGATCAAG2513823RGTGACTGATATCCCATCCACATTCTC26243FCATCAAGTATTCTTGGGCGGAGATA25141243RCCATTTCTAGACATAAATGTTGACGCTATT30244FTTCACCTTTAGAGTGACGCACTTG24153244RTACTACTGGCCAACGACGTTTCG23246FCAACTACAAAATGGTCTTAGAACTACAC28107246RCATCTGCAAAACATGCACCTGATATAC27247FGGCATGAATGTATTAAGGTCTACAATC27149247RGACATGGTTCGTAACACGGAAAT23248FGTAATCGGATTGGCCCATTG2086248RGGCACTGAACAGAAGATTGCAG22250FCCTATCAAATAAATACTTGGCCCACA2684250RGAGCCCAGATAGGCAAAGATCA2225FAGACAGAGGAAGACCGATGCA2118925RGACGGCAGGGAGGTTGAT18260FAAAAAACTTACCTCTGTGCGCATTTA2691260RCTGCTCTAGCCAATCTCGTACCAC24262FCTACTTCATGATGCTTTGCACGA2389262RACGACGAACAGAACGCTGATGAT23263FTCTGATACGAATCAAATGCACGAT24116263RGATTAGAATTTGTGGTCTGCCTCAG25264FTCATTGACCAGCCAATCTTGC21105264RTGTGCAAGTAAATTACAAGAGCGAAC26265FAAATTAAGCCGAGTATGAACTGATTG26103265RCTAGGGGCAACCGTGTTAAGTG22267FGCCATAGTGATGGTGGCACATAG23122267RCGTCTGAAATCGGATGTCTAGGAAT25268FCCACATACTATTCATAACTAAAGCGTTG28101268RGTTTCAAATTGGTGCGTGGAT21269FCTTAGATTGAAGATTGCTACACATAGATT29120269RGTATTCTTCATCCAAGAGTATATTGAGGT29271FTGTGTAAACCGGTTGGGATCTTC23163271RGGAACGTGGTACTCTACGTCTAGTTTG27272FATGTAGTGGAGGATCAACCCATGAT25157272RTAAACTGCCCTCTGGCTTGCGACA24274FATATCGTAGATATGCCTCTGCTCTGG2686274RCTTTCCTATGCTTCAAATGATACGACT27275FATAGTACTGAAATTCTTTGTCTGTGTAGC29110275RTGTGTAGTGAACAAATTCTCGCATAATC28276FAGGGGCACAAAGAGGTTGCTCTA23143276RTTATGTCCGATTGAGAAATCCAACTTC27278FCGGCGACATCCACTCTTTACA21145278RCATCTGTATCGTTCTCTGATGAATTGG27280FTGCAAATGCACTCCTACGGTACTC24127280RGGCTTGCTGGATGCAACTGT20281FTTTCTGGACCATACCGACCACTG23151281RAAGCTTGATATCACATAGAGGCACAT26282FGATGATTATCCCAAGATTGCCTTG2497282RCACCTCGCGTCATACAACGTCT22283FTTGCTACAATATTTGCGGAGGAAG24124283RGTAATGGACTAGTTTATCAATTCCTACTG29284FCCTGTCCATTCGAACCCAATATG23128284RAGACGGGAAGTGGGCTGTAAATAG24285FAAGTGCGTGGTAATGAGATCCTG23194285RGAAACTCAAGCTGCAACTCAGTCA24288FCTGAAATTCTCCAATAGGCTGTTGAT26167288RACGTTACCACATCAGCAAAATACCAT26289FGGCTCCTTACAAATCATGGCAAC23162289RACCTCAGCACTCTTGACAGCATC2328FTAGTGCCATATGTGTATTGTACCGTTC2712028RCAATCATAACTTACTATCAAGATGAACTG29292FCAACGTTGCACTTGCAGGAGT21149292RACGACGGACGGTGACTCAAG20293FGAGGACAAAGGAAACCCATACGTG24191293RGTCTATCCAACTCGATCTCGTGTCA25295FTTGGTATTAGGAAAAGGCACATAGTTC2770295RTGTGCCTGGGAATATGGTTATGTC24296FGAGGATCCACAAATTGCAGCTCT23115296RACTATCCAAGAGCATACGCACGAC24297FAAGGCATACTGTGTACAACTCCCAAT26160297RGGCTAGAGGATGCTACATCTTTGGTG26299FGGCAATAAATTAAGCTCTGGGTGT24205299RGGTGAGAAGACAAACCTAATCATCA252FGTAGGAGATCAATAACGACGAGACAAA271622RCCATGACGACGCAATAGTTGGTAC24300FCTCGTCCTGGACTTGGAGGTAG22121300RATGCTGATACCCAAATATGAACTCCA26301FATGAACGTTGACGTCGGATGC21125301RGAGAGGTTCTAGAGTTGCTTTTCATCA27304FCCTTCCAAACGTAGAGGCCATACT24128304RAGTTAGGGATCGAAATAGGACATCTG26307FAGTGGTTTTCAAGTTTGGACCTCA24117307RAGGTACCCTGGGTCCATATCAC22308FTTACCCACAACTAGTGCCACAACA24110308RGATATCAATGAGTGTGCTGCCAAAC25309FGTTGCATGTGAAGTCATTGCTTATG25170309RAGCTCTAATAAAAATAGCGGCAAAGC26310FGGCGTTGAATTAGTGATAGGGATTG25139310RCCTCTATCTGATATTTGCCCAACAAC26311FATACTTGCATATACGTGGAACCAGAG26167311RCAAAGAGCCGATCAGCCAGT20312FGGACAGATGCCGGGCTAAGAT21113312RGAACCTGATAACCTAATAACGGTGGA26313FTAGAGAAGGGTTTTTTTCTTGGTAACT27158313RCACATTAGTGACGGTTCAAGATGTA25314FGACCGACGGTGATGAGAAACAG22117314RCCAATCCTGGTGTTCCAGTAGGAG24315FTGGTACTCCAACACACCACTGATAAT26160315RGTCGTTTTGATCAAGTTGGCTCAG24318FCATCGTAGGTAGATGATCTCAACTTTT27165318RTTGGTGTGTTGCCCAATTCAG2131FACGAGTTCCAGGTGATGCAGA2115931RGCAGCTGAACGAGCTGTTCCTA22320FAACTCTGAAAATGACCAGGTGCTAAC26111320RGCTTTGGTAGAGGAATCATTGGTG24321FTTCAGAACTTCAGCGTATATGCACCT26138321RCACCTTTAGTCAAGCTGACCCTGTAAC27322FCTTTATTGGCAGCGACGACCTA2270322RCCCGTGCTAGCTGTCGAGGT20323FGACACGCCGGTCACAGAGATA21105323RCAGGTAAATGCCATCACCAGAGTC24325FTCCATCACTTGATCCCAACGAG2283325RCCTTCTAAGCAGACACAGATTCTACTA27326FCAAGAGAGTTACAAAACCGACTTTACA27144326RGTTGTTGTTGCTGTAGGGGCTAA23327FCTGTGTGCTGGTTTTATCCGTTTATAG27144327RACACGCTGTGCATCAATCAAGTG23328FATCGCCATCCATCAGACACG20120328RCGTGCGACAGACGTGTCCTA2032FCTTGTGCTGATTGCTGCTGCTA2214732RCTTTGTTTAATTCACACGATATCATT2634FTGTGGCGGTGTGGGTAGCA198934RGCTGCTTGGTGCGGAGCTT1936FCTTTGACTTGGGCATCGAACT2110036RTCGCTCCCTCCACACTACG1937FGCCAAATTCCGGTATACGCA2013937RTTTTGTCCACGTTATATTTTCCTCG2539FCAAAAATTGACACACCTTATTTACAGG2714139RGAAAAATTATCAGTTCATGAGGGGAC2640FCCTCTGTGTGTTTCTTCCAAAATACC2616640RCCATGTACCAAATTTACATCGTTTCA2642FTTGGGGCGGTCGATTCTCG1911142RAACGGAGCAAGCTATCACTCACC2343FGACAAGGGACATAGTTGAGCCAAA2414343RGTGCTCTTATAATTTGGACAGTTCTGTTAC3045FCAGAAACCACACTGTTCTAGACACGA2610845RCGAGTTCTCTGGAGTTGCTGG2146FCACTACAAATATCATTCTGCTGAAGAC2713046RGTCAGGAGATTTGAAAGCAATATAGATG2848FCGAAACACTTGGCTCTATTATTGAACTTAC3012648RGAACAACATGCAGCATCCAAAGTA2449FAAAACGACATTTAAAAAGGGACCAG2512749RTTATTAAGTTATTACTATTACCGTGGCTA294FGGAACCTTTTCGGATTGTCAG212984RTGCAGGTAACGTGATGAGTAAAACTG2650FTGGTTTATGCATTAGTTACATATTTGG2710350RGAGTTCAACAGTTAATAAACAACATACAGA3052FGGCACAAGGAAGGCAAGAGTG219152RTTCACCTGTTCGCCTTCCTGTA2253FCATGCTGGAACACACAGGAAGATA2415953RTAGGACATCCCCTAACCAGTAGTCTAT2755FTTTTAGAAATCACATCTTATATTATGGAAC3013155RTCTGGAGCAAGTTTGTAGAATTGTACTT2859FCCCATTCTGCTACACTCTTCTCCATAC2714959RGATTCCTCAGTTGGCTATAGTTGGC2560FCTGTGAAGTCCCTGTGTTCCAAAC2415860RATGCCCAAAGCGGCTATTAC2062FCAACGTCTTGTTGGTGAACGA2117962RCTCAAGATAAGGAATGAATGTCAAAGA2763FGGGAAAAAAACTCAGATCTAGCTTACC2711463RAACTTAGAGAAATTTGACTCTGGATGC2765FACCCAGCGATTTAAGCAACAATTC248965RCAACTCACACTGAAGCAAGTGTTTT2566FTGGGGGAAAGAACAAGAAAGTTTG2411366RTCCTAACAGTAAGTTGTTTCTCATCC266FCTGGCGCAGGTAAAACCCTA20886RGTAGCGCCGTCGTTCAAGA1970FATGTCCCATTGTAGAGCGCTGAT2317370RAGCAAATGCAGATGATTTGAAGAA2471FGATGACATGTAGATTAGTTGAAGAGATT2813671RATTCACATGGTTGCTATGCTACTC2476FGATGAAATTGCCTTAGCCTCATATTTC2712976RGTCTTTGCTATTTGGTTAGTACTAGGTC2877FCTTAGCTTGCGCTACCGTGT2012477RCCATCTCTTAACTTTTATTATTGATTTCC2979FGAACAAGTTATTTGATTCTGACCCAT2611479RGATAAGGTGCTTACTCTTGCTGGT2482FTAATCCGACAATGGATGACACC227282RTGGCGCTGCTCCTTGTAAC1984FCGGTGTCTATCCTAGCGCTTCA2214284RAGGCATTTGATGTACATGTGCTG2385FTGAGTCTCCCGTGAGCAGCAG219985RCGCCATCGGGACCATTTGA1986FGATGATTTGGTTGCACACTTGA228586RGTGAAAATAATTCGCTACAAACCTTC2687FCCAACCACGTCATCGACCTC208087RGTCCTGGCAGGGGATATTGC2088FCCCTAGCCGTTGATTTGGTG2012988RCATGTGTCCTAAGATAGTTGGATGG2589FATGAGCGAACCAAGTCGACAAT2210089RGCAATAGAACCTGCGAGCTGTG228FCTAACAAGGGTAGGAATCGAGAGAAA261488RAACAGAATTCAGAAAAAGGTTGTCAC2690FTGTACCAGGCATTTGCATCTCTG239090RTCGCAGATCCAAGCCTGAAAATAC2491FGGTTATGGTCTGAAGCGGTCATT2312791RGTGTTCCATGATTATGAATTTTTTAGC2792FGATATTTTGGCTTTGTTAGATATACTG2715192RCACAAATCCTTATATTGTACTCCCTCAAT2995FGCACTCCTTTCGCTGAGCATAC2217895RGGGGACCCTTCACTATTCATTCA2396FTGCTGCACCATCAAAATCAGC219896RTGCATGCCTGCGGCTACTG19連接引物信息見(jiàn)表D.2。連接引物信息引物名稱(chēng)序列(5'--3')長(zhǎng)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