Cell line Authentication kit

The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - Mouse aortic endothelial cells (MAECs)

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - Peripheral blood mononuclear cells (PBMC)

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - Peripheral blood mononuclear cells (PBMC)

Get tips on using TACS® 2 TdT Fluorescein Kit to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using ApoAlert™ DNA Fragmentation Assay Kit to perform TUNEL assay cell type - Islets of langerhans (Beta cells)

Get tips on using ApoAlert™ DNA Fragmentation Assay Kit to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using STEMdiff™ Definitive Endoderm Kit to perform Stem cell Differentiation media Differentiation of Human hESCs into Endoderm

Get tips on using MethylFlash Hydroxymethylated DNA Quantification Kit to perform DNA methylation profiling Whole genome profiling - human germ cell cancer

Stem cells have the unique ability to self-renew or differentiate themselves into various cell types in response to appropriate signals. These cells are especially important for tissue repair, regeneration, replacement, or in the case of hematopoietic stem cells (HSCs) to differentiate into various myeloid populations. Appropriate signals refer to the growth factor supplements or cytokines that mediate differentiation of various stem cells into the required differentiated form. For instance, HSCs can be differentiated into dendritic cells (with IL-4 and GM-CSF), macrophages (with m-CSF) and MDSCs (with IL-6 and GM-CSF). Human pluripotent stem cells (hPSCs) and induced pluripotent stem cells (iPSCs) can be first cultured in neural differentiation media (GSK3𝛃-i, TGF𝛃-i, AMPK-i, hLIF) to form neural rosettes, which can be differentiated into neural or glial progenitors (finally differentiated into oligodendrocytes). Neural progenitors can be finally differentiated into glutaminergic (dibytyryl cAMP, ascorbic acid) and dopaminergic (SHH, FGF-8, BDNF, GDNF, TGF-𝛃3) neurons. Thus, it is important to first identify the self-renewing cell line: its source and its final differentiation state, followed by the supplements and cytokines required for the differentiation, and final use. Timelines are another thing that is considered. For instance, it takes 7-10 days to form neural rosettes from iPSCs and 3 days to differentiate neural progenitors to neurons. Finally, the stability for stem cell culture media varies. It is advised to make fresh media every time when differentiating HSCs to myeloid populations, whereas neural differentiation media may remain stable for two weeks when stored in dark between 2-8C.