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Cholerny Spammer
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 Posted: Thu 6:08, 24 Feb 2011 Post subject: new balance outlet Angelica ISSR-PCR reaction syst |
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Angelica ISSR-PCR reaction system and optimization
Reaction polymorphic bands disappear. Different primers, annealing temperature are not the same, and thus determine the appropriate annealing temperature is necessary. Rm primers used in this experiment a 8 ~ C ~ Tm +12 ~ C range to 864 primers [(ATG) 6, Tm = 48 ~ C], for example, automatically generated by the PCR amplification temperature of 8 to select the best annealing temperature. Figure obtained from the 864.48-l} 1 into the evil school of Medicine in 2008 to approximately 2 = fj31 Lane Ta primer is 48 ~ C. 1-7: Bow l Ren concentration 0.1.0.2.0.3. 0.4.0.5,0.6.0.8 ~ mol / L Figure 3 lead Ren concentration of PCR reaction 1-8: Wen Ha is: 40.0. 41.3-43.8.47.3. 52.1. 55.9. 58.4.60. O ring 4, PCR annealing temperature on the reaction of 3 Conclusions ISSR molecular marker technology is based on PCR, the amplified RAPD marker bands, although more stable, but is also affected by changes in reaction conditions and the impact of different species. The experimental results show that different ISSR amplification reaction system will have a greater impact, thus affecting the accuracy of ISSR analysis,[link widoczny dla zalogowanych], and accuracy and stability of ISSR band spectrum is the genetic diversity of the premise. In order to take advantage of this molecular marker genetic diversity of Angelica, access to high repeatability and reliability of ISSR band spectrum, it is necessary to impact factors were selected and optimized to establish the most appropriate iSSR reaction system. In this study, Angelica DNA were used to establish a reproducible, high resolution of ISSR reaction system, ie 25L reaction system, the template DNA concentration of l0 ~ 30ng, 2.5L's 10XPCRbuffer (Mg ¨ free),[link widoczny dla zalogowanych], MgC122. 0mmol / L, dNTPs0.2mmol / L, Taq enzyme 1. Ou,[link widoczny dla zalogowanych], O. primer 6p, mol / L; amplification program: 94 ~ C denaturation 5min; 94 ℃ denaturation 3Os, primer annealing at the optimal screening temperature annealing 45s, 72 ℃ extension of 2min, a total of 39 cycles,[link widoczny dla zalogowanych], after the end of 72 cycles ℃ for 5min. Using this system, the 100 ISSR primers screened, Article l0 response has been clear bands,[link widoczny dla zalogowanych], polymorphic primer on Angelica ISSR genetic diversity research will be reported.
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[link widoczny dla zalogowanych]
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