FokI NEB#R0109

Restriction Enzymes FokI

Experiment
Restriction Enzymes FokI
Product
FokI NEB#R0109 from New England BioLabs
Manufacturer
New England BioLabs

Protocol tips

Protocol tips
After PCR amplification, samples are subjected to restriction enzyme digestion with FokI and BtsCI and desalting using Oasis purification plates, followed by analysis by MALDI-TOF MS.

Publication protocol

HPV DNA was amplified with PGMY09/11, composed of two nondegenerate pools of L1 consensus primers37. For the second round, we used nested PCR primer pairs; a sense primer PV-rfmpF specific to bases 6584–6603 (5′-GCMCAGGGHCAYAAGGATGAATGG-3′; M = A/C, H = A/C/T, Y = C/T) and an antisense primer PV-rfmpR specific to bases 6657–6626 (5′-GTACTDCKDGTRGTATCHACMACGGATGTAACAAA-3′; D = A/G/T, K = G/T, R = A/G). As illustrated in Figure 1, sequences underlined in each primer were modified to insert FokI restriction recognition site in the PCR products. Nucleotide sequence positions were numbered according to HPV genotype 16 (GenBank accession number AY686584).

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Manufacturer protocol

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