3261

急需徵求翻譯達人的人......一篇論文

One of the milestones of this project is the generationof a comprehensive repository of full-length genescoding for all human peptidases. From the analysis ofPuente et al. (2003)

the final number of functionalpeptidases in the human genome is 460. To set upthe conditions for their automated PCR amplification

we initially selected for a pilot study 23 genes rangingfrom 384 to 3261 bp coding for peptidases and wescreened different DNA polymerases

PCR conditionsand cDNA sources in 96 MTP format. Fig. 1 showsthat the degree of detectable PCR amplifications of thereactions assembled by the robotic system varies considerablydepending on the polymerase used and onthe cDNA template. Strikingly

cDNA retrotranscribedfrom a mixture of total RNA extracted from 55 differenthuman tissues supported a lower amplificationoutcome in respect to cDNA from testis. However

it isevident (Fig. 1B

“combined” percentage) that the successrate can be increased by enlarging the diversity ofthe template cDNAs.The vast majority of the experimental steps in thecloning phase were performed in 96 MTP format

including PCR reaction set up

PCR products purification

DNA ligation

E. coli transformation

PCR-basedscreening for positive clones

amplification of the bacterialclones

plasmid DNA purification and preparationof glycerol cell banks. The overall success rateof the high-throughput cloning is shown in Fig. 2.As expected

longer genes are subjected to a higherhindrance

especially in the PCR amplification and inpreserving a 100% sequence identity. In a single-roundstreamlined procedure

the percentage of the positiveclones ranged between 39 and 55% for genes longer orshorter than 1100 bp

respectively. However

it shouldbe underlined that extremely strict criteria were appliedfor the evaluation of the sequence identity.
這個項目的當中一個里程碑是世代全長基因一間全面貯藏庫編制程式為所有人的peptidases 。

從分析Puente 等。

(2003)

最後數字的功能peptidases 在人的染色體是460 。

設定條件為他們自動化的PCR 放大作用

我們最初地選擇了為一中間試驗23 基因排列從384 到3261 bp 編制程式為peptidases 和我們被篩選的不同的DNA polymerases

PCR 情況並且DNA 來源在96 MTP 格式。

無花果。

1 展示程度可發現的PCR 放大作用反應由機器人系統裝配可觀地變化根據polymerase 被使用和DNA 模板。

醒目

DNA retrotranscribed從總核糖核酸混合物從55 被提取不同人的組織支持了更低的放大作用結果關於DNA 從睪丸。

但是

它是顯然(圖 1B