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Reagent

ul/rxn

rxns

ul needed (1.5x)

10x T4 Buffer

1.15

600

690

437

5M NaCl

0.12

600

7245.6

1 mg/ml BSA

0.6

600

360

228

H2O

0.73

600

438277.4

MseI (enzyme)

0.12

600

7245.6

EcoR1 (enzyme)

0.28

600

168

106.4

Total

3

600

18001140

Add 6 ul template to each plate with Benchsmart. Cover and seal each plate, vortex, centrifuge and incubate at 37C for 8 hours. Followed by 65C in EcoBGC oven for 1 hour.

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