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,2016,36(7):1464-1469
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
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犃狀犗狆狋犻犿犻狕犲犱犆犺犾狅狉狅狆犾犪狊狋犐狊狅犾犪狋犻狅狀犪狀犱犆犺犾狅狉狅狆犾犪狊狋犇犖犃
犈狓狋狉犪犮狋犻狅狀犘狉狅狋狅犮狅犾犳狅狉犆犺犻狀犲狊犲犠犻犾犱犌狉犪狆犲狊
XIEHaikun,JIAOJian,FANXiucai,ZHANGYing,JIANGJianfu,
SUNHaisheng,LIUChonghuai
(ZhengzhouFruitResearchInstitute,ChineseAcademyofAgriculturalSciences,Zhengzhou450009,China)
犃犫狊狋狉犪犮狋:Matureleavescolectedfrom犞犻狋犻狊犱犪狏犻犱犻犻,犞.犪犿狌狉犲狀狊犻狊,犞.犺犲狔狀犲犪狀犪and犞.犮犺狌狀犵犪狀犲狀狊犻狊
wereusedforchloroplastisolationandcpDNAextractioninthisstudy.Thetwomethodswerethecolumn
plantchloroplastDNAoutandmodifiedhighsaltlowpHmethod,andtheresultswerecomparedwitheach
other.(1)BothmethodshadseparatedthechloroplastofChinesewildgrapes,butthemodifiedhighsalt
lowpHmethodobtainedhigherconcentrationandlessimpurityofchloroplastthanthatofcolumnplant
chloroplastDNAout.SothemodifiedhighsaltlowpHmethodwasmoresuitableforchloroplastisolation.
(2)ThevalueofOD260/OD280ofcpDNAextractedbythecolumnplantchloroplastDNAoutwasbetween
1.28and1.36,andtheconcentrationwasbetween4.2ng·μL
-1and7.8ng·μL
-1,whichdidnotmeet
thedemandofsubsequentchloroplastgenomesequencing.Incontrast,thevalueofOD260/OD280ofcpDNA
extractedbythemodifiedhighsaltlowpHmethodwasbetween1.84and1.90andtheconcentrationwas
between2514.4ng·μL
-1and4133.7ng·μL
-1,sothecpDNAextractedinthiswaywasextremely
highqualityandpure.Asaresult,thecpDNAextractedbythemodifiedhighsaltlowpH methodmeet
thedemandofsubsequentchloroplastgenomesequencing.Asaconclusion,themodifiedhighsaltlowpH
methodisolatedintactchloroplastandextracthighqualitycpDNAofChinesewildgrapessimplyand
quickly.AndthecpDNAmeetthedemandofsubsequentchloroplastgenomesequencing.Itwasalsoa
criticalsteptomakefurtherresearchofchloroplastgenomesof犞犻狋犻狊L.
犓犲狔狑狅狉犱狊:Chinesewildgrapes;chloroplastisolation;cpDNA extraction;columnplantchloroplast
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Table1 UVscanningresultsofcpDNAextractedbytwomethods
?U
Species
Method OD260
/OD280 OD260/OD230
R
Concentration/(ng·μL-1)
hPQ
犞.犱犪狏犻犱犻犻
Ⅰ 1.30 0.58 7.8
Ⅱ 1.84 2.10 3708.5
iPQ
犞.犪犿狌狉犲狀狊犻狊
Ⅰ 1.35 0.67 5.6
Ⅱ 1.90 2.14 2876.0
jkPQ
犞.犺犲狔狀犲犪狀犪
Ⅰ 1.36 0.51 4.2
Ⅱ 1.87 2.07 2514.4
mnPQ
犞.犮犺狌狀犵犪狀犲狀狊犻狊
Ⅰ 1.28 0.44 4.8
Ⅱ 1.89 2.13 4133.7
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Note:ⅠandⅡstandforthecolumnplantchloroplastDNAoutandthemodifiedhighsaltlowpHmethod
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modifiedhighsaltlowpHmethod(Ⅱ)
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:
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