RestrictionEnzymSigma-Aldrich 代謝組學克隆和表達
【簡單介紹】
品牌 | Sigma-Aldrich | 貨號 | Restriction Enzymes 克隆和表達 |
---|---|---|---|
規(guī)格 | 基因組編輯工具 | 供貨周期 | 一周 |
主要用途 | 代謝組學 |
繼ZFN(Zinc Finger Nucleases)技術后,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、更經濟的方式實現基因組特定位點的編輯。
【詳細說明】
基因組編輯工具-CRISPR/Cas9 | ||
繼ZFN(Zinc Finger Nucleases)技術后,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、更經濟的方式實現基因組特定位點的編輯。憑借過去10年在基因組編輯領域的豐富經驗積累以及專業(yè)的生物信息學平臺,Merck已經成功設計出覆蓋人類,小鼠和大鼠三個物種的所有基因的CRISPR/Cas9載體,并可以提供在線定制服務,以及完整的CRISPR實驗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,以幫助科學家實現基因功能的快速篩選、規(guī)?;哪P徒⒁约八幬镒饔煤Y選等。
- 高效:優(yōu)化的載體設計,大限度提高轉染效率,簡化篩選工作
- 特異:特殊的gRNA設計和雙切口酶系統,大限度提高特異性
- 全面:產品齊全,可提供質粒、RNA、慢病毒載體、RNP等形式,涵蓋人、大鼠、小鼠、植物等多個物種,更有Sanger Arrayed和Broad Pools全基因組文庫以及重要通路的亞文庫
- 掌控:強大的慢病毒全基因組文庫可輕松進行高通量篩選,全面掌控人或小鼠的基因組
CRISPR實驗--載體構建和表達 |
• CRISPR/Cas gRNA設計和選擇 • crRNA定制合成 • 克隆和表達 • 抗生素 • GenElute™質粒DNA快速提取kit • PCR擴增和核酸電泳 • SP6/T7 體外轉錄試劑盒 • GenElute™總RNA提取試劑盒 • 慢病毒包裝系統 • 培養(yǎng)皿/離心管/槍頭等基礎耗材 |
Molecular BiologyCloning & Expression | ||
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.
Buffer | |||||
Restriction Enzyme | SA (B7531) | SB (B8781) | SL (B3782) | SM (B3158) | SH (B3657) |
Acc I (R6142) | 100 | 0-10 | 10-25 | 1-10 | 0-10 |
Alu I (R6885) | 100 | 50-75 | 25-50 | 25-50 | 0-10 |
Apa I (R4258) | 100 | 10-25 | 50-75 | 50-75 | 0-10 |
Bam HI (R0260) | 100 | 100 | 75-100 | 100 | 25-50 |
Bcl I (R8631) | 100 | 100 | 25-50 | 100 | 100 |
Bgl II (R6377) | 100 | 100 | 25-50 | 100 | 100 |
Bln I (R3131) | 25-50 | 50-75 | 0-10 | 25-50 | 100 |
Bsm I (R3635) | 0-10 | 50-75 | 0-10 | 25-50 | 100 |
Cfo I (R1761) | 75-100 | 50-75 | 100 | 50-75 | 25-50 |
Cla I (R7763) | 100 | 100 | 75-100 | 100 | 100 |
Dde I (R4256) | 50-75 | 75-100 | 25-50 | 25-50 | 100 |
Dpn I (R8381) | 100 | 75-100 | 50-75 | 75-100 | 75-100 |
Dra I (R4381) | 100 | 75-100 | 100 | 100 | 50-75 |
Ecl XI (R3884) | 25-50 | 100 | 25-50 | 25-50 | 50-75 |
Eco RI (R6265) | 100 | 100 | 25-50 | 50-75 | 100 |
Eco RV (R2756) | 25-50 | 100 | 0-10 | 25-50 | 50-75 |
Hae III (R5628) | 50-75 | 50-75 | 75-100 | 100 | 25-50 |
Hind III (R1137) | 50-75 | 100 | 25-50 | 100 | 50-75 |
Hpa I (R8507) | 100 | 25-50 | 25-50 | 50-75 | 25-50 |
Kpn I (R1258) | 75-100 | 10-25 | 100 | 25-50 | 0-10 |
Nco I (R8761) | 50-75 | 50-75 | 50-75 | 50-75 | 100 |
Nde I (R5509) | 25-50 | 100 | 10-25 | 50-75 | 100 |
Nhe I (R5634) | 100 | 25-50 | 100 | 100 | 10-25 |
Not I (R8506) | 10-25 | 50-75 | 0-10 | 25-50 | 100 |
Nsi I (R5884) | 50-75 | 100 | 10-25 | 50-75 | 100 |
Pst I (R7023) | 25-50 | 25-50 | 10-25 | 25-50 | 100 |
Pvu I (R1508) | 50-75 | 75-100 | 25-50 | 50-75 | 100 |
Pvu II (R2631) | 25-50 | 25-50 | 25-50 | 100 | 25-50 |
Rsa I (R4756) | 100 | 50-75 | 100 | 50-75 | 0-10 |
Sac I (R5268) | 100 | 0-10 | 100 | 50-75 | 0-10 |
Sal I (R0754) | 0-10 | 25-50 | 0-10 | 10-25 | 100 |
Sau 3A (R0762) | 100 | 25-50 | 25-50 | 75-100 | 0-10 |
Sca I (R5007) | 0-10 | 100 | 0-10 | 75-100 | 100 |
Sfi I (R8256) | 25-50 | 25-50 | 75-100 | 100 | 25-50 |
Sma I (R4503) | 100 | 0-10 | 0-10 | 0-10 | 0-10 |
Spe I (R5257) | 75-100 | 75-100 | 75-100 | 100 | 100 |
Sph I (R7135) | 50-75 | 75-100 | 25-50 | 100 | 75-100 |
Ssp I (R6759) | 75-100 | 75-100 | 10-25 | 75-100 | 100 |
Taq I (R9507) | 50-75 | 100 | 25-50 | 50-75 | 50-75 |
Xba I (R7260) | 100 | 75-100 | 75-100 | 75-100 | 100 |
Xho I (R6759) | 25-50 | 75-100 | 10-25 | 25-50 | 100 |
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).
Final concentration (mM) | |||||
SA (B7531) | SB (B8781) | SL (B3782) | SM (B3158) | SH (B3657) | |
Tris-acetate | 33 | ||||
Tris-HCl | 10 | 10 | 10 | 50 | |
Mg-acetate | 10 | ||||
MgCl2 | 5 | 10 | 10 | 10 | |
K-acetate | 66 | ||||
NaCl | 100 | 50 | 100 | ||
Dithioerythritol (DTE) | 1 | 1 | 1 | ||
Dithiothreitol (DTT) | 0.5 | ||||
2-Mercaptoethanol (BME) | 1 | ||||
pH at 37°C | 7.9 | 8.0 | 7.5 | 7.5 | 7.5 |
Categories
- Buffers
- Digestions
- Enzyme activity
- PAGE
- Solid phase extractions
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