繼ZFN（Zinc Finger Nucleases）技術(shù)后，Merck在2013年推出新一代基因組編輯工具--CRISPR/Cas9，讓研究人員以更快、更經(jīng)濟(jì)的方式實(shí)現(xiàn)基因組特定位點(diǎn)的編輯。憑借過去10年在基因組編輯領(lǐng)域的豐富經(jīng)驗(yàn)積累以及專業(yè)的生物信息學(xué)平臺(tái)，Merck已經(jīng)成功設(shè)計(jì)出覆蓋人類，小鼠和大鼠三個(gè)物種的所有基因的CRISPR/Cas9載體，并可以提供在線定制服務(wù)，以及完整的CRISPR實(shí)驗(yàn)workflow解決方案。此外，默克與Sanger Institute合作開發(fā)了人、小鼠全基因組CRISPR 文庫(kù)，以幫助科學(xué)家實(shí)現(xiàn)基因功能的快速篩選、規(guī)模化的模型建立以及藥物作用篩選等。

• 高效：優(yōu)化的載體設(shè)計(jì)，大限度提高轉(zhuǎn)染效率，簡(jiǎn)化篩選工作
• 特異：特殊的gRNA設(shè)計(jì)和雙切口酶系統(tǒng)，大限度提高特異性
• 全面：產(chǎn)品齊全，可提供質(zhì)粒、RNA、慢病毒載體、RNP等形式，涵蓋人、大鼠、小鼠、植物等多個(gè)物種，更有Sanger Arrayed和Broad Pools全基因組文庫(kù)以及重要通路的亞文庫(kù)
• 掌控：強(qiáng)大的慢病毒全基因組文庫(kù)可輕松進(jìn)行高通量篩選，全面掌控人或小鼠的基因組

CRISPR實(shí)驗(yàn)--載體構(gòu)建和表達(dá)

• CRISPR/Cas gRNA設(shè)計(jì)和選擇 • crRNA定制合成 • 克隆和表達(dá) • 抗生素 • GenElute™質(zhì)粒DNA快速提取kit • PCR擴(kuò)增和核酸電泳 • SP6/T7 體外轉(zhuǎn)錄試劑盒 • GenElute™總RNA提取試劑盒 • 慢病毒包裝系統(tǒng) • 培養(yǎng)皿/離心管/槍頭等基礎(chǔ)耗材

The aim of molecular cloning is to insert the gene-of-interest (GOI) into a plasmid vector. This vector is then inserted into a cell that will express the protein encoded by the GOI. Once protein is expressed, the protein function can be studied as it affects the cell signaling, morphology or other aspects. Alternatively, the protein can be expressed in large quantities that can then be studied directly with other techniques. We have a wide variety of products for use across the cloning & expression workflow.

expression workflow.

Cloning Products

• SnapFast™ Vectors • Cloning Enzymes • Restriction Enzymes • Biomatrica™ Nucleic Acid Stabilization • Custom DNA Oligos

Restriction Enzyme Buffer Reference

Sigma^® Life Science’s restriction enzyme collection has been optimized for digestion using five unique buffers. Reference below the enzyme activity in each buffer (table 1), and the preferred buffer for each enzyme (table 2).

Table 1
Activity (%) of each Sigma restriction enzyme in each Sigma buffer. The preferred buffer for each enzyme is highlighted in bold.
 

Table 2
The complete formulation of each Sigma Restriction Enzyme Buffer. The formulation of the storage buffer for each Sigma restriction enzyme is given on the catalog page (see links in table 1).
 

Categories

• Buffers
• Digestions
• Enzyme activity
• PAGE
• Solid phase extractions