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The low number of hits may be due to the fact that Gal/GalNAc lectin is a glycoprotein which is more difficult to digest using the common digestion protocol.
Total Kjeldahl nitrogen (TKN) was analysed using the Kjeldahl method, ammoniacal nitrogen (NH4-N) using KCl extraction, total phosphorous (TP) and available phosphorus (AP) (by acid digest) using the stannous chloride method.
Total DNA was determined from agarase/papain digest using the Hoescht 33258 method.
This amplification was repeated on 10 μl of mung bean nuclease digest using the same N-1A and NTA-1A primers, but with a 20-cycle PCR.
To generate blunt ended DNA fragments suitable for cloning into pHORF3 [ 28] a restriction enzyme digest using the blunt end cutting enzymes AfeI, AluI and CviKI-1 (NEB, Frankfurt, Germany) was performed.
The in silico experiments suggested that a complete genome digest using the restriction enzyme StuI should produce approximately 31,000 fragments in the 1.5 2 kb size class (representing 2.7% of the genome) and provide >20× sequencing coverage.
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The two pre-weighed samples were digested using the same digestion procedure enumerated above.
Spots were excised from the gel and transferred into Microtiter plates, destained and digested using the Ettan Digester (GE Healthcare).
Samples were digested using the microwave oven digester (BERGHOF Speed Wave Four).
Lymph nodes were removed and mammary glands were digested using the Miltenyi Fatty Tissue Digestion Kit.
The emulsion was digested using the standardised in vitro digestion protocol recommended by the Infogest COST Action (Minekus et al., 2014), involving 2 h gastric followed by 2 h of small intestinal simulation.
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