基因组编辑工具-CRISPR/Cas9
继ZFN（Zinc Finger Nucleases）技术后，Merck在2013年推出新一代基因组编辑工具--CRISPR/Cas9，让研究人员以更快、更经济的方式实现基因组特定位点的编辑。

基因组编辑工具-CRISPR/Cas9

继ZFN（Zinc Finger Nucleases）技术后，Merck在2013年推出新一代基因组编辑工具--CRISPR/Cas9，让研究人员以更快、更经济的方式实现基因组特定位点的编辑。凭借过去10年在基因组编辑领域的丰富经验积累以及专业的生物信息学平台，Merck已经成功设计出覆盖人类，小鼠和大鼠三个物种的所有基因的CRISPR/Cas9载体，并可以提供在线定制服务，以及完整的CRISPR实验workflow解决方案。此外，默克与Sanger Institute合作开发了人、小鼠全基因组CRISPR 文库，以帮助科学家实现基因功能的快速筛选、规模化的模型建立以及药物作用筛选等。

• 高效：优化的载体设计，大限度提高转染效率，简化筛选工作
• 特异：特殊的gRNA设计和双切口酶系统，大限度提高特异性
• 全面：产品齐全，可提供质粒、RNA、慢病毒载体、RNP等形式，涵盖人、大鼠、小鼠、植物等多个物种，更有Sanger Arrayed和Broad Pools全基因组文库以及重要通路的亚文库
• 掌控：强大的慢病毒全基因组文库可轻松进行高通量筛选，全面掌控人或小鼠的基因组

 

CRISPR实验--载体构建和表达

• CRISPR/Cas gRNA设计和选择 • crRNA定制合成 • 克隆和表达 • 抗生素 • GenElute™质粒DNA快速提取kit • PCR扩增和核酸电泳 • SP6/T7 体外转录试剂盒 • GenElute™总RNA提取试剂盒 • 慢病毒包装系统 • 培养皿/离心管/枪头等基础耗材

Molecular Biology Cloning & 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