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- 第一步,计算质粒的分子量。假设一个DNA碱基对(bp)的平均重量为650道尔顿(Da),一摩尔的碱基对(bp)的重量为650克,即任何双链DNA模板的分子量可以通过取其长度(bp)与每个碱基对重量的乘积来估计。
First, calculate the molecular weight of the plasmid. Assuming that the average weight of a DNA base pair (bp) is 650 Daltons (Da) and that one mole of a bp weighs 650 g, the molecular weight of any double-stranded DNA template can be estimated by taking the product of its length (in bp) and weight per base pair.
计算公式:Plasmid molecular weight [g/mol] = plasmid size [bp] x 650 [Da/bp] - 计算每微升中质粒的摩尔数。
Next, calculate the number of moles of plasmid per microliter.
计算公式:Moles of plasmid per microliter = plasmid concentration [g/µL] / plasmid molecular weight [g/mol] - 然后,用阿伏伽德罗数(6.022 x 1023molecules/mole)计算每微升的质粒分子数。
Then, calculate the number of plasmid molecules per microliter, using Avogadro’s number (6.022 x 1023molecules/mole).
计算公式:Molecules of plasmid per microliter = moles of plasmid per microliter [moles/µL] x Avogadro’s number [molecules/mole] - 稀释质粒原液(分子/微升),得到100微升的溶液,所需浓度为1 x 109个分子((vg)每微升)。
Finally, dilute the plasmid stock (molecules/µL) to obtain a 100 µL solution with the desired concentration of 1 x 109 molecules (vector genomes (vg) per microliter).
计算公式:Plasmid stock (100 µL) = (desired concentration [molecules/µL] x 100 µL) / plasmid molecules [molecules/µL]
- 第一步,计算质粒的分子量。假设一个DNA碱基对(bp)的平均重量为650道尔顿(Da),一摩尔的碱基对(bp)的重量为650克,即任何双链DNA模板的分子量可以通过取其长度(bp)与每个碱基对重量的乘积来估计。
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