Supply of T4 DNA ligase 100000units/pack (NEB) M0202. Manufactured by M/s.NEW ENGLAND BIOLABS (NEB) multiple suppliers as per attached annexure
2
Phusion High-Fidelity PCR Master Mix with HF buffer (NEB) F531S
3
Monarch® PCR & DNA Cleanup Kit (5 µg) For purification of up to 5 µg of concentrated, high-quality DNA from PCR and other enzymatic reactions. The kit should utilize a bind/wash/elute workflow with minimal incubation and spin times. Eluted DNA should be ready for use in restriction digests, DNA sequencing, ligation and other enzymatic manipulations. Should allow purification of ssDNA oligonucleotides (= 18 nt) and dsDNA fragments (= 15 bp) . The columns should have zero retention of buffer and no carryover of buffers. Typical yields must be in the following range: DNA (50 bp to 10 kb): 70–90%DNA (11–23 kb): 50–70%ssDNA =18 nt and dsDNA = 15 bp: 70-85%, (NEB) T1030S
4
Nco-I HF (NEB)Catalog no. R3193L, 20000units/ml5000 units
NotI-HF (NEB) R3189L restriction enzyme. Enzyme must be able to digest DNA in 5-15 minutes and provide flexibility to digest overnight without degradation. 20000units/ml, 2500units
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