MCB Laboratory Research Protocols

Details Included

Purpose

Required Materials

Recipes

Protocols

Special Notes on Procedures

Waste Disposal Procedures

Special Precautions Needed

References

Cell Culture Protocols

MCBLP-001CC: Culturing of Human Mesenchymal Stem Cells/hMSCs
This is to culture and start the initial seeding of Human Mesenchymal Stem Cells (hMSCs), which are adherent fibroblast -like cells isolated from bone marrow, adipose, and other tissues.
MCBLP-002CC: Subculturing and expansion of Human Mesenchymal Stem Cells/hMSCs
MCBLP-003CC: Cell Counting with a Hemocytometer
MCBLP-004CC: Thawing Cryopreserved Cells
MCBLP-005CC: Cryopreservation of Cells
MCBLP-006CC: Generation of Growth Curve (for Cell Lines)
MCBLP-007CC: Isolation and Culture of Human Gingival Fibroblasts (HGnF)
MCBLP-008CC: Alkaline phosphatase (ALP) Staining
MCBLP-009CC: Modified Alkaline Phosphatase (ALP) Quantification Assay
MCBLP-010CC: Nile Red Quantification for Lipids
MCBLP-011CC: Oil Red O Staining

Gene & Protein Purification, Expression and Analysis Protocols

MCBLP-001GP: Agarose Gel Electrophoresis for Separation of DNA samples
The purpose of agarose electrophoresis is to separate DNA based on size, as well as to visualize it after amplification or DNA extraction (e.g. such as plasmid or genomic DNA extraction). DNA can then be extracted from the gel for further analysis and processing.
MCBLP-002GP: PCR using Platinum® PCR SuperMix by Invitrogen
MCBLP-003GP: PureLink® Genomic DNA Extraction by Invitrogen™
MCBLP-004GP: SDS-PAGE Gel Preparation (for Proteins)
MCBLP-005GP: SDS-PAGE for Protein Separation
MCBLP-006GP: SDS-PAGE Staining and Destaining
MCBLP-007GP: WESTERN BLOTTING Gel Blot Transfer to Membrane (for Proteins)
MCBLP-008GP: Real-Time PCR (qPCR)
MCBLP-009GP: Nucleic acid quantification using spectrophotometric method
MCBLP-010GP: Protein quantification using spectrophotometric method
MCBLP-011GP: Polyacrylamide Gel Preparation (for Nucleic acids)
MCBLP-012GP: Polyacrylamide Gel Electrophoresis (PAGE) for Nucleic Acids
MCBLP-013GP: Gel Staining (PAGE for Nucleic Acids )

Immunohistochemical Studies Protocols

MCBLP-001IH: Tissue Preparation for Histological Bone Samples
This is to preserve tissue samples permanently in as life-like state as possible and to allow for the preparation of thin, stained sections for histological analysis.

Microbiological Studies and Analysis

MCBLP-001MC: Bacterial Growth Curve Using 96-Well Plate
This is to measure the growth of bacteria using a 96-well plate instead of the classic cuvette method at OD600. The protocol only requires 100 μL of media to obtain the growth curve.
MCBLP-002MC: Bacterial Biofilm Formation Assay using a 96-Well Plate