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The PCR products were digested under the conditions specified by the enzyme supplier (New England Biolabs Inc, Beverly, MA, USA).
All four primers were included in a 10 μl reaction with 20 100 ng genomic DNA, HotMaster Taq (Eppendorf™), 0.2 mM each dNTP and the enzyme supplier's buffer.
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Up-scaling of these different enzyme processes has predominantly been done in close collaboration between enzyme suppliers and early adopters in the industry, very often in collaboration with engineering companies.
We also tested commercial cellulase mixtures from other enzyme suppliers.
'On-site' enzyme production is economically advantageous and efficient [ 26] and it reduces the cost and supply dependency on commercial enzyme suppliers.
For qPCR using chemically-modified hot-start enzyme AmpliTaq Gold, a cognate buffer containing 10 mM Tris pH 8.0, 50 mM KCl and 2.5 mM MgCl2 is usually recommended by the enzyme suppliers.
By leveraging well-proven bisulfite technology and the rapid progress of enzyme suppliers, CLE Sugar can achieve a high yield of total biomass carbohydrate conversion to monomeric lignocellulosic sugars.
For Southern hybridization, approximately, 4-5 μg of buffalo, cattle, goat and sheep genomic DNA were subjected to restriction digestion using RsaI enzyme following supplier's (NEB) specifications.
Approximately, 5 μg of buffalo genomic DNA from both the sexes was digested with RsaI restriction enzyme following supplier's (NEB) specification.
10 μL of PCR products was digested with 0.3 unit of Hha1 enzyme according to the supplier's recommended procedure (Biolabs New England).
Enzyme activity reported by the supplier was 100 units/g dry materials.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com