19 Activity 5. Digestion of genomic DNA with 3 different restriction endonucleases Assemble all reaction components on i

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19 Activity 5 Digestion Of Genomic Dna With 3 Different Restriction Endonucleases Assemble All Reaction Components On I 1 (64.5 KiB) Viewed 71 times

19 Activity 5. Digestion of genomic DNA with 3 different restriction endonucleases Assemble all reaction components on ice, add the restriction endonuclease last Table 3 Restriction endonuclease digest of genomic DNA Volume in final reaction Tube 1 Tube 2 Tube 3 Details Reaction components Amount in final reaction BamHI Hindtil EcoR1 gDNA BamHI 150ng 5 units 5 units Hindill 5 units EcoR1 Restriction endonuclease buffer 25 ng/ 20 units/ 20 units/l 20 units/ Provided as 10x stock Add to make final reaction volume 20ul Up to 12 hours, 37°C 1x in final reaction Add to make final reaction Water volume 20 DNA digestion Dilution of enzymes: BamHI can be diluted in Diluent Buffer A: Hindill can be diluted in Diluent Buffer B: EcoR1 can be diluted in Diluent Buffer C. Enzymes should be diluted just prior to use.