All the reagents and chemicals were of high purity and analytical grade. Standard paclitaxel was procured from MP Biomedicals (USA). Media components used for the growth and maintenance of taxol-producing endophytes were purchased from Hi-Media (Mumbai). The electrophoresis reagents, molecular grade chemicals used for DNA isolation were purchased from SDFCL (India) and Sigma-Aldrich (USA), respectively. Pre-coated Silica gel 60, F254 TLC plates, and HPLC solvents were of HPLC grade and procured from Merck (Germany). Universal primers for ITS and dbat genes were procured from Sigma (USA) and Bioservice, respectively.
Pre coated silica gel 60
Manufactured by Merck Group
Sourced in Germany
Pre-coated Silica gel 60 is a laboratory product used for chromatographic separation and purification processes. It consists of silica gel particles with a specific pore size and surface area, pre-coated with a stationary phase to facilitate selective separation of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
F254 tlc plate, by Merck Group (1 mentions)
Hplc solvents, by Merck Group (1 mentions)
Avance 300 nmr spectrometer, by Bruker (1 mentions)
Ascend 400 ft nmr spectrometer, by Bruker (1 mentions)
Ft ir spectrum 400 spectrophotometer, by PerkinElmer (1 mentions)
Microtof q 2 mass spectrometer, by Bruker (1 mentions)
J 810 spectropolarimeter, by Jasco (1 mentions)
Silica gel 60, by GE Healthcare (1 mentions)
Uv 1800 spectrophotometer, by Shimadzu (1 mentions)
4 protocols using pre coated silica gel 60
1
Paclitaxel production by endophytes
2
Identification and Characterization of Fatty Acids
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All solvents used in this study were available from Italmar Co., Ltd. (Bangkok, Thailand). All commercial fatty acids were purchased from Sigma-Aldrich (Singapore). The structure and purity of all identified compounds were confirmed by FT-NMR, and GC-MS analyses. 1H and 13C NMR spectra were provided by a Bruker AVANCE 300 NMR spectrometer (Bruker BioSpin AG, Fällanden, Switzerland), operating at 300 MHz and 75.5 MHz, respectively. FCC and quick column chromatography (QCC) were conducted by using Merck silica gel 60 (<0.063 mm) and Scharlau GE 0048 silica gel 60 (0.02–0.06 mm), respectively. For TLC, MERCK precoated silica gel 60 F254 plates were used. Spots on TLC were visualized under UV light and sprayed with an anisaldehyde-sulfuric acid reagent, then heated.
3
Characterization of Natural Compounds
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Optical rotations were measured on a JASCO-1020 polarimeter. Electronic Circular Dichroism (ECD) was recorded on a JASCO J-810 spectropolarimeter. UV spectra were collected on a Shimadzu UV 1800 spectrophotometer. IR spectra were recorded in the ATR mode using a PerkinElmer FT-IR Spectrum 400 spectrophotometer. 1H and 13C NMR spectra were recorded on a Bruker ASCEND 400 FT-NMR spectrometer, operating at 400 MHz (1H) and 100 (13C) MHz. HR-ESI-TOF-MS spectra were measured with a Bruker micrOTOF-QII mass spectrometer. Unless otherwise indicated, column chromatography was carried out using Merck silica gel 60 (particle sizes less than 0.063 mm) and GE Healthcare Sephadex LH-20. For thin-layer chromatography (TLC), Merck pre-coated silica gel 60 F254 plates were used. Spots on TLC were detected under UV light and by spraying with anisaldehyde-H2SO4 reagent followed by heating.
4
Lipid Class Separation and Quantification
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In addition to total fat content and FA composition analyses, total extracted lipids (TL) from fish liver were separated into phospholipids (PL) and triacylglycerols (TAG) classes using thin layer chromatography (TLC) as described earlier by Pickova et al. (1997) (link) and Mráz & Pickova (2009) (link). TLC plates (20 cm × 20 cm; pre-coated Silica gel 60; 0.20-mm thickness from Merck, Darmstadt, Germany) were selected as stationary phase and solvents (hexane, diethylether, and acetic acid; 85:15:1, v/v/v) were added to TLC chamber. PL and TAG sample areas were identified by comparison with a reference standard separately revealed in iodine. Areas of interest were scraped off the TLC plates. The PL class was extracted successively with chloroform:methanol (2:1, 2 ml, v:v); chloroform:methanol (1:1, 3 ml, v:v) and chloroform (2 ml), while TAG was extracted three times in succession with chloroform (2 ml). Samples were evaporated under nitrogen gas, recovered in 0.5 ml hexane, and stored at −20°C until further analysis.
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