For Blue/White Cloning of a DNA Fragment
Blue/white screening is one of the ways scientists determine if cloning has been successful. In this kit, students complete a ligation, a transformation, and an assay, which is based upon the blue/white colony screening protocol. If the gene that expresses β-galactosidase is interrupted by the cloning plasmid, then white colonies will grow because the enzyme is not able to cleave the X-Gal indicator. If the β-galactosidase gene is not cut by the plasmid, then the X-Gal is cleaved into galactose and a blue indigo compound. So, if the colonies are blue, then the insertion failed. Comprehensive Background section explains each step in the process. Three convenient halt points are indicated in the procedure so the experiment can be performed over several days in three 60–70 minute sections.
This Kit includes the DNA fragment, linear plasmid, T4 DNA Ligase, competent cells, X-Gal, and all the necessary reagents. An incubator, two waterbaths, a shaking incubator or shaking waterbath, microwave or hot plate, automatic micropipet and tips, spectrophotometer, balance, centrifuge, and microcentrifuge are all required and available separately.
Materials Provided in Kit |
Storage upon Receipt |
Host Strain |
-20°C |
Vector |
-20°C |
Insert |
-20°C |
T4 DNA Ligase |
-20°C |
Ligase Assay Buffer |
-20°C |
Control Plasmid |
-20°C |
Transformation Solutions |
2°C – 8°C |
Media Components |
RT |
Antibiotic |
-20°C |
All solutions should be kept tightly sealed and stored at above define temperature. These reagents should remain stable for at least 1 year in their unopened containers.
Items | Cat. # | Size |
---|---|---|
Blue White Screening Kit | SCMT009 | 10 Preps. |
Blue- White Screening Kit | SCMT009A | 20 Preps. |
For Blue/White Cloning of a DNA Fragment
Items | Cat. # | Size |
---|---|---|
Blue White Screening Kit | SCMT009 | 10 Preps. |
Blue- White Screening Kit | SCMT009A | 20 Preps. |
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