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- W2275939185 abstract "With genomic DNA of Torreya grandis ‘Merrillii’,an optimization experiment was performed in terms of Taq DNA polymerase,Mg2+,DNA template,deoxynucleotide triphosphates(dNTP)s,and primer concentration.The optimized system was as follows: a total polymerase chain reaction(PCR) volume of 20 μL included 30 ng DNA,16.67 nkat Taq polymerase,0.350 μmol.L-1 primer,1.625 mmol.L-1 Mg2+,0.250 mmol.L-1 dNTPs,and 2 μL 10 × PCR buffer.Then,a PCR amplification program was established with denaturing at 94 °C for 5.0 min followed by 35 cycles of denaturing at 94 °C for 30 s,annealing for 45 s(annealing temperatures varied with primers) with extension at 72 °C for 1.5 min,extension at 72 °C for 7 min,and finally maintained at 4 °C.Based on this optimized system,34 inter simple sequence repeat(ISSR) primers suitable for the species were screened from 77 primers with gradient PCR being used to pro-pose the optimal annealing temperature.This experimental result has laid a foundation for genetic map con-struction in T.grandis ‘Merrillii’." @default.
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- W2275939185 date "2010-01-01" @default.
- W2275939185 modified "2023-09-26" @default.
- W2275939185 title "Optimization of an ISSR-PCR system and primer screening for Torreya grandis ‘Merrillii’" @default.
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