The present work focuses on the value addition of cotton stalks through xylan extraction and enzymatic production of xylooligosaccharides (XOS); a pentose based prebiotic. Chemical analysis revealed that cotton stalk contains neutral detergent fibre 87.66%, acid detergent fibre 72.81%, hemicellulose 14.85%, cellulose 53.08%, and acid detergent lignin 19.73%. The xylan yield from cotton stalks was ranged from 1.75 to 7.36% when recovered with lithium hydroxide, while it reached up to 14% by using sodium and potassium hydroxides. The maximum recovery of xylan was attained at 4% NaOH together with steam and 8% KOH under overnight incubation. Commercial xylanase enzyme was used for enzymatic hydrolysis of xylan to produce XOS. The XOS concentration was ranged from 0.83 to maximum value of 2.19 mg/ml in the hydrolysate at pH 5.0, enzyme dose of 5 U, for 4 h of hydrolysis time. The findings substantiated the potentiality of cotton stalk as a raw material for xylan extraction vis a vis production of XOS.