Materials And Methods Of Gram Negative Bacteria Escherichia Coli

823 Words Jan 7th, 2015 4 Pages
2. Materials and Methods:
The keratin was extracted from Chicken feather which were collected from the local market of Raipur (21.14°N 81.38°E) Chhattisgarh, India. The 91% food grade alginate (sodium salt form) was purchased from Loba Chemie. The β-mercatpoethanol (mol. wt. 78.13) from LOBA Chemie, sodium dodecyl sulfate (SDS) (mol. wt. 288.38) and urea (mol. wt. 60.06) was purchased from Merck, and ethanol was purchased from Sigma-Aldrich. Antimicrobial strain of gram negative bacteria Escherichia coli (E. coli) [MTCC 118] and gram positive bacteria Bacillus subtilis (B. subtilis) [MTCC 121] was purchased from MTCC Chandigarh, India.

2.1. Fabrication of Porous Scaffold from Keratin/Alginate binary blend:
The extraction of keratin from Chicken feather was done by using method, described by Yamauchi et al. 14. The homogenized stock solution of keratin 7% (w/v) and alginate 4% (w/v) were prepared and store at ambient temperature. The keratin and alginate samples were characterized by FTIR before using it as blend material. The blend solution was prepared using the optimum parameter, obtained by the CCD model of RSM. The freeze-extraction method as described by the Wang et al. 12, were used to fabricate the scaffold.

2.2. Compatibility study of keratin and alginate:
The compatibility/miscibility between polymer of keratin and alginate was confirmed using “X-ray Diffractometer” (XRD) and “Fourier Transform Infrared Spectroscopy” (FTIR). The XRD (X’pert PRO, PANalytical)…

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