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Protocols

protocols
Protocol

Optimized Protocol to Create Deletion in Adherent Cell Lines Using CRISPR/Cas9 System

Experiment Tags: Single Cell, Genomics, Molecular Biology, Gene Expression, CRISPR

Experiment Summary: In this article, we described an improved HTGTS (iHTGTS) method, in which size-selection beads were used to enhance reaction efficiency and a new primer system was designed to be compatible with Illumina Hiseq sequencing. iHTGTS is a powerful method for comprehensively assessing Cas9 off-target effect.

protocols
Protocol

Protocol for Rate-Controlled Cryopreservation and Thawing of Mammalian Cells

Experiment Tags: Biological Techniques, Cryopreservation, Cooling, Freezing

Experiment Summary: In this protocol, we describe the general procedure for cryopreservation of cultured cells.

protocols
Protocol

Protocol for Mitochondria Transfer between Mesenchymal Stem Cells and Glioblastoma Stem Cells

Experiment Tags: Mitochondria Transfer, Mesenchymal Stem Cells, Glioblastoma Stem Cells, Cell Interaction, Cell Biology.

Experiment Summary: This protocol facilitates the transfer of mitochondria from mesenchymal stem cells (MSCs) to glioblastoma stem cells (GSCs). This protocol allows for the investigation of the effects of mitochondria on GSC metabolism, proliferation, and response to therapy by isolating mitochondria from MSCs and transferring them to GSCs.

protocols
Protocol

Intraductal Injection of Adenoviral Cre for Temporally Controlled Breast Cancer Modeling in Mice

Experiment Tags: Breast Cancer, Mouse Model, Intraductal Injection, Adenoviral Cre, Temporal Control

Experiment Summary: This protocol outlines a technique for injecting adenoviral Cre (Ad-Cre) intraductally into mice mammary glands to create animal models of breast cancer. By employing cell-type-specific promoters to target mammary epithelial cells (MECs), this strategy enables fine temporal control of cancer induction and cell-type specificity. The process is described in the protocol, which includes creating and caring for floxed mice, monitoring the growth of breast tumors, and providing postoperative care.

protocols
Protocol

3D Tumorsphere Culture Protocol for Cancer Research

Experiment Tags: 3D Tumorsphere Culture, Cancer Research, Matrigel Embedding, Tumor Tissue Culture, Cell Transduction, Cryopreservation, Cell Counting

Experiment Summary: This protocol outlines a comprehensive procedure for 3D tumorsphere cultivation, including cell transduction, 3D tumorsphere culture using Matrigel, primary tumorsphere culture from fresh tumor tissue, spheroid passaging, and cryopreservation. The protocol aims to replicate the 3D environment of solid tumors, aiding researchers in studying tumor biology and conducting drug screening.