**MBI Fermentas Endonuclease**
When it comes to cloning, time is of the essence. You need to excise your insert from a vector using two different restriction enzymes—but checking your old enzyme poster, you realize they don’t work in the same buffer. That means another hour added to your schedule for sequential digestions. What if you could simplify the process?
Introducing **Thermo Scientific FastDigest Restriction Enzymes**, designed to streamline your workflow with one universal buffer that works for all 176 FastDigest enzymes. With this powerful solution, you can enjoy:
- **100% activity** in a single buffer, compatible with modifying enzymes
- **Complete digestion** in just 5–15 minutes
- No star activity—thanks to optimized incubation times and enzyme amounts
- **FastDigest Green Buffer** allows direct loading on gels, no extra loading dye needed
- Tested protocols for plasmid DNA, PCR products, and genomic DNA
Say goodbye to double digest stress and hello to more time for other experiments.
**Blue-White Screening**
This technique helps identify successful clones by using a lacZα fragment. When an insert disrupts this sequence, colonies turn white on X-gal plates, indicating successful insertion. However, false positives can occur. To avoid this, use **positive selection vectors**, which express a lethal gene that is only inactivated when a DNA insert is present. This ensures only recombinant cells survive.
**Restriction Digest for Insert Verification**
To confirm the presence of your insert, perform a restriction digest. Isolate plasmid DNA using a kit like the Thermo Scientific GeneJET Plasmid Miniprep Kit. Use our **REsearch restriction site mapping tool** to plan your digestion. Run the digested sample on an agarose gel to check for expected band sizes.
**Colony PCR**
Another way to verify inserts is through colony PCR. Use insert-specific or a combination of vector- and insert-specific primers. This method works well for inserts under 3 kb. Simply pick a colony, run PCR, and analyze the results.
**Sequencing**
For the most accurate confirmation, sequencing is the gold standard. Isolate plasmid DNA, then perform Sanger sequencing using vector-based primers. Sequencing across the entire insert ensures the correct sequence is present.
Whether you're working with endonucleases, proteins, or PCR, MBI Fermentas offers reliable tools for your research. Visit our website to learn more about **FastDigest Restriction Enzymes** and see how easy and efficient they can make your cloning process.
**For Research Use Only. Not for Diagnostic Procedures.**
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