recombinant dna bacterial transformation student instructions plasmid digest
DESCRIPTION
Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest. Recombinant Transformation Plasmid Digest. Obtain four 1.5 ml reagent tubes labeled: - pAmp - pKan - 2XRB”( Restriction Buffer ) - B/H” (BamHI and HindIII enzymes). - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest](https://reader035.vdocuments.mx/reader035/viewer/2022062802/56814627550346895db336fb/html5/thumbnails/1.jpg)
Recombinant DNA Bacterial Transformation
Student InstructionsPlasmid Digest
![Page 2: Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest](https://reader035.vdocuments.mx/reader035/viewer/2022062802/56814627550346895db336fb/html5/thumbnails/2.jpg)
Recombinant TransformationPlasmid Digest
Obtain four 1.5 ml reagent tubes labeled: - pAmp - pKan - 2XRB”(Restriction Buffer) - B/H” (BamHI and HindIII
enzymes).
![Page 3: Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest](https://reader035.vdocuments.mx/reader035/viewer/2022062802/56814627550346895db336fb/html5/thumbnails/3.jpg)
Use the matrix below as a checklist while adding reagents to the pAmp and pKan tube.
Use a fresh pipette tip for each reagent. Add all reagents directly to the solution.
Recombinant TransformationPlasmid Digest
NOTE: The buffer is always added before the enzymes.
Tube 2X RBB/H
__________________________________________________________ pAmp 7.5 µl
2 µl
pKan 7.5 µl2 µl
![Page 4: Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest](https://reader035.vdocuments.mx/reader035/viewer/2022062802/56814627550346895db336fb/html5/thumbnails/4.jpg)
Recombinant TransformationPlasmid Digest
Place the reaction tubes in a 37oC water bath and incubate for 30-60 minutes.
![Page 5: Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest](https://reader035.vdocuments.mx/reader035/viewer/2022062802/56814627550346895db336fb/html5/thumbnails/5.jpg)
Recombinant TransformationPlasmid Digest
After incubation, freeze the reaction at -20oC until you are ready to proceed to Ligation. You can store reactions at this temperature for several days.