Page 31 - nai udaan december

Materials and Methods

• Tea samples (Processed tea, Green tea, Seabuckthorn tea, Fresh tea leaves)
• Reagents (Folin-ciocalteu reagent, Sodium carbonate solution)
• Equipment (Spectrophotometer, Table top Centrifuge)
• Various lab consumables (Falcon tube, Eppendorf tube, Whatman filter paper

Preparation of tea extract

Weigh tea sample (1 gm), and add 5 ml 70% methanol in a falcon tube.
Incubate at 65 oC for 10 min, vortex, and spin the samples at
6000 rpm for 10 min.
Collect the supernatant, re-spin & store the
supernatant for
further use at 40C.

(B) Optimizing the paper disc exper-
iment
Bacterial colony inoculated into 5 ml
sterile nutrient broth in a sterile test tube

Tubes were subjected to gentle shaking and incubated at 370 C for 10-12 hours

1. Plated different dilutions of
bacterial culture in plates- (2 OD, 0.6
OD, 0.1 OD) (~200µl)
2. Incubation temperatures (370C,
280C)
3. Different amount of tea extracts
(10µl, 50µl, 100µl)

Plates incubated for 16-24 hours and zoneof inhibition recorded

fnlacj -2023 xÉ<Ç =cÉxÉ 29