Samples of the dairy beverage were stirred for 1 min on a vortex stirrer (VX-200, Labnet
), and then centrifuged for 20 min at 5000 rpm and 12[grados]C.
En este proceso los ensayos se llevaron a cabo en tubos falcon de 50 ml los cuales se llenaron con 35 g de muestra y se procedio a introducirlos en un bano seco AccuBlock modelo D1200 (Labnet
, USA) para controlar la temperatura del proceso.
Ambarees, also known as "Labnet
al-Jarra," due to its storage in jars, was once a common dairy product made in most households of the Bekaa Valley.
Inicialmente se sensibilizo la placas con 100 [micro]L/pocillo de la proteina recombinante MSP5r a 2 [micro]g/mL, diluido en buffer de carbonato-bicarbonato 50 mM (pH 9,6) y se incubo la placa a 4 [grados]C en camara humeda (Labnet
, 311DS, EUA) por toda la noche.
About 0.5 [micro]L of CNF suspension was dripped using a Labnet
micropipette onto a 300-mesh carbon coated formvar copper grid, and the water was allowed to evaporate.
10 [micro]L of the PCR products was then separated on 2% agarose gel (Biopioneer Co., USA) thereafter in 1x TAE running buffer (Biopioneer Co., USA) by electrophoresis at 80 volts (Labnet
International, Power station 300) using 2 [micro]L of blue/orange DNA loading dye (6x) (Promega Co., USA) and stained with 0.5 [micro]g/mL ethidium bromide (Life Technologies Co., USA).
As part of manual and semiautomated processes, the reaction vial was heated using an Accublock Digital Drybath heater block (Labnet
, Woodbridge, New Jersey, USA) with a modified aluminium adaptor.
Phase separation was performed by centrifugation (Spectrafuge 6C Compact model, Labnet
Touchdown PCR was performed on a MultiGene[TM] thermocycler (Labnet
International, Inc., USA) using the following cycling conditions: denaturation at 94[degrees]C for 1 minute, followed by a hybridization step using a 0.5[degrees]C decrease for each cycle between 65 and 55[degrees]C, with 10 additional cycles at 55[degrees]C, and the extension step at 72[degrees]C for 3 minutes.
Electrophoresis was carried out at 100 V for 30 minutes using Endoro[TM] Gel XL, Labnet
The amplification of the reverse-transcription products was carried out by PCR (1 [micro]l of cDNA, 1.5 mM Mg[Cl.sub.2], 20 [micro]M dNTPs, 10 pmol of primers (Promega, Charbonnieres, France) (Table 1) and 2.5 units of recombinant Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA) in a MultiGene cycler (Labnet
, Woodbridge, NJ).
Se tomaron entre 50 a 100 mg de tejido conservado, el tejido se lavo con agua destilada tres veces agitando con Vortex (Labnet