Gas chromatograph attached with MIDI Sherlock Microbial Identification System and PLFA Analysis software

Gas chromatograph attached with MIDI Sherlock Microbial Identification System and PLFA Software is being used for identification of bacterial strains based on gas chromatographic (GC) analysis of extracted microbial fatty acid methyl esters (FAMEs) and FAME profile compared with profile stored with MIDI’s Sherlock bacterial library and PLFA in environmental soil samples quantified by using Sherlock’s PLFA analysis software.

Instrument Details


Make:    Agilent Technologies
Model:    Gas chromatograph 7890A attached with MIDI’ s Sherlock Microbial Identification and PLFA Analysis software

Detailed Specifications:
  • The GC has two inlets but currently dedicated only for MIDI and PLFA software’s and have FID detector with electronic pneumatics control.
  • The GC has detector electronics and has full-range digital data path for peaks quantifications.
  • Has full electronic pneumatics control (EPC) with auxiliary devices for inlets and detectors and has pressure set points and control precision to 0.001 psi
  • Has EPC with capillary columns provides four column flow control modes
  • Installed column (capillary) for fatty acids (FAME) and has fully integrated Automatic Liquid Sampling (ALS) system with time programming and display for initiation of on/off, method start, etc.,
  • Operating temperature range is approximately +4 °C to 350 °C with set point resolution of 1°C.
  • Should have carrier and makeup gas settings selectable for H2, N2, and O2 gases with compatible gas regulators and cylinders of standard/ISI make.
  • System has more than 10 fast ramps with split/split less inlet compatible for FAME capillary column
  • The GC should have fully equipped with compatible identification software’s like MIDI and other window based software’s along with latest configuration PC and laser printer
  • Have compatible UPS (5KVA) for continuous power back

Available Accessories:
GC with FID detector, MIDI’s Sherlock Microbial Identification Software, PLFA software, two stage regulator with control panel to regulate gases, gas cylinders (Oxygen, Nitrogen and Hydrogen), 24- place Vacuum manifold
Contact:  Dr M.P Sharma
Principal Scientist (Microbiology)
Ph: 0731-2437971(O), 9926012261(M)
Email: mahaveer620@gmail.com, dsrdirector@gmail.com





About Instrument:
Agilent make GC (7890A) is attached with MIDI’ Sherlock Microbial Identification and PLFA Analysis software. The GC system has FID detector with ultra2 capillary fatty acid column along with oxygen, nitrogen and hydrogen gases fixed with control panel with two stage regulators for flame and carrier gases. The MIDI Sherlock Microbial Identification System is a fully automated gas chromatographic analytical system that identifies over 3,000 microbial species based on their unique cellular fatty acid/FAME profiles stored in library (RTSBA6 Rapid/TSBA6 Standard) and PLFA absolute yield (ng/g soil) of environmental and soil samples is quantified from PLFA extracted samples.

Working Principal

GC Microbial fatty acid profiles are unique from one species to another, and this has allowed for the creation of very large microbial libraries. After extraction of fatty acids methyl esters (FAME) from bacterial cultures samples analysed on gas chromatograph using FID detector and peak naming is done through MIDI’s Sherlock bacterial library (RTSBA6) and identified based on fatty acids comparison pattern of strains stored in the library software. The PLFA quantification in environmental soil samples is done after extraction and elution of desired phospholipid fractions (PLFA, NLFA) by employing details protocols involving extraction, separation and elution of phospholipid fractions through column chromatography and transesterified samples analysed on GC autosampler using Sherlock’s PLFA analysis software and absolute yield of PLFA is quantified after data transformation.

Applications

Gas chromatograph 7890A attached with MIDI’s Sherlock Microbial Identification is being used to characterize microbial cultures and PLFA Analysis software is used for quantifying absolute yield of phospholipid fractions in environmental and soil samples.

User Instruction

1. FAME extracted samples (using GC/HPLC grade solvents) stored in screw capped PTFE GC 2 ml vials can be analysed for FAME and identified through comparison of FAME pattern stored in the MIDI’s Sherlock’s library
2. Environmental and soil samples with extracted PLFA fractions (Fatty acids separated through column chromatography and uses GC/HPLC grade chemicals and solvents) can be analysed for PLFA quantification