I’m thinking of reducing steps for insert production.
ATM I have following steps:
2. agarose gel with PCR products
3. gel extraction of insert of interest
4. restriction reaction
5. enzyme removal <- I often use KpnI/BamHI and can’t go with heat inactivation
6. ligation with backbone and transformation
I know that restriction can be performed on agarose slices but never done it myself. This would save a bit of time and reagents, plus avoid DNA losses in step 5.
Anyone got first-hand experience with this? If ‘yes’, link to a protocol would be greatly appreciated.