???? ???????(PCR) ????????DNA(RAPD)

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???? ???????(PCR) ????????DNA(RAPD)
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• ?????PCR????????
• ??RAPD???,????RAPD???????

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• ???????(polymerase chain reaction,PCR)??????DNA???
  ?????????DNA?????????????????????,???????????????3
  ???,??????,DNA??2n??????2530????DNA???106109??

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• ???????DNA(Random Amplification Polymorphic
  DNA)????PCR?????????????,??????????????????????,??
  ???????DNA????,????????????????????????,?EB???????
  ??????????????RAPD??,?????DNA??????????,??????????
  ????????PCR?????,???????????????????,????3???????
  ??????,???????DNA??,????????????DNA??????????,????
  ?????????????????????,??PCR????????,????????,????P
  CR?????????????????????????

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• ?????DNA??

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• PCR?,PCR????,???,??,???????,?????4?dNTP,??,Taq???
  ,Buffer,MgCl2,DNA Marker,????

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• 1???????
• ???????DNA????,???5?RAPD??????PCR?????PCR???20ul??
  ???

??? ???? PCR???? ???
??DNA(??) 100 ng/ul 100 ng 1.0 ul
???Buffer 10 1 2.0 ul
4?dNTP??? 10 mmol/L 500 umol/L 1.4 ul
Mg Cl2 25 mmol/L 1.4 mmol/L 1.2 ul
Taq DNA??? 5unit/ul 1unit 0.2 ul
??Primer(??) 10 umol/L 0.5 umol/L 1.0 ul
????? 12.2 ul
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• 2?PCR??
• ???PCR???PCR????????,????93????4min93???4min35
  ???1min72???2min,?45???72???10min?
• 3???
• ??7 ul PCR???2 ul????????,???1.2
  ??????,5v/cm??1h?,?????????????,??????
• 4??????????
• ?1(?) ?0(?) ????,???????????????Jacard?????,??
  ??i ?j ??????????(F),???D1-F, ?SPSS????????,?????
  ????,???????????????

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• ??????,???????????????(F),????????????
• ???????????,??????????????

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? 2 ???RAPD???10?????????????? Tab.2 Ten valid arbitrary primer, its sequence and PCR results in RAPD of Camellia japonica L. ? 2 ???RAPD???10?????????????? Tab.2 Ten valid arbitrary primer, its sequence and PCR results in RAPD of Camellia japonica L. ? 2 ???RAPD???10?????????????? Tab.2 Ten valid arbitrary primer, its sequence and PCR results in RAPD of Camellia japonica L. ? 2 ???RAPD???10?????????????? Tab.2 Ten valid arbitrary primer, its sequence and PCR results in RAPD of Camellia japonica L. ? 2 ???RAPD???10?????????????? Tab.2 Ten valid arbitrary primer, its sequence and PCR results in RAPD of Camellia japonica L.
? ? Primer ? ? Sequence ????? Number of bands ?????? Number of polymorphic bands ??????() Percentage of polymorphic bands
S1 GTTTCGCTCC 15 8 53.3
S2 TGATCCCTGG 6 2 33.3
S4 GGACTGGAGT 8 5 62.5
S6 TGCTCTGCCC 9 3 33.3
S7 GGTGACGCAG 9 2 22.2
S10 CTGCTGGGAC 6 3 50.0
S11 GTAGACCCGT 5 2 40.0
S12 CCTTGACGCA 6 5 83.3
S184 CACCCCCTTG 8 7 87.5
S185 TTTGGGGCCT 6 4 66.7
Total 78 41 52.6
Average 7.8 4.1 52.6
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? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L. ? 4 ??????????????? Tab.3 Genetic distant matrix of six-variety Camellia japonica L.
? ? Number ? ? Variety 1 2 3 4 5 6
1 American Dahong
2 Fendan 0.2500
3 China-America Hong 0.0909 0.2143
4 Feizhua Furong 0.1579 0.2414 0.1754
5 Yudan 0.2381 0.0841 0.2000 0.2844
6 Hua Feizhua Furong 0.1927 0.2613 0.2294 0.0796 0.2692
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