miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Contamination can affect cell characteristics, i.e., growth, metabolism, and morphology leading to unreliable and erroneous experimental data. Depending on the source of contaminants, one can detect contamination by using a light microscope, gram stain, isothermal amplification, or PCR. Bacteria and fungi can usually be identified by optical microscopy. Mycoplasma in cell cultures cannot be detected visually. Hence, these microbes can go unnoticed for long periods and are determined using dedicated assays. Early and rapid identification of contaminants is vital to detect, handle and prevent contamination for good cell-culture practices. However, detection and identification can be challenging and tricky based on usual visual identifications. Hence it is essential to use a standard contamination detection kit to detect and maintain best practices.
Contamination can affect cell characteristics, i.e., growth, metabolism, and morphology leading to unreliable and erroneous experimental data. Depending on the source of contaminants, one can detect contamination by using a light microscope, gram stain, isothermal amplification, or PCR. Bacteria and fungi can usually be identified by optical microscopy. Mycoplasma in cell cultures cannot be detected visually. Hence, these microbes can go unnoticed for long periods and are determined using dedicated assays. Early and rapid identification of contaminants is vital to detect, handle and prevent contamination for good cell-culture practices. However, detection and identification can be challenging and tricky based on usual visual identifications. Hence it is essential to use a standard contamination detection kit to detect and maintain best practices.
Contamination can affect cell characteristics, i.e., growth, metabolism, and morphology leading to unreliable and erroneous experimental data. Depending on the source of contaminants, one can detect contamination by using a light microscope, gram stain, isothermal amplification, or PCR. Bacteria and fungi can usually be identified by optical microscopy. Mycoplasma in cell cultures cannot be detected visually. Hence, these microbes can go unnoticed for long periods and are determined using dedicated assays. Early and rapid identification of contaminants is vital to detect, handle and prevent contamination for good cell-culture practices. However, detection and identification can be challenging and tricky based on usual visual identifications. Hence it is essential to use a standard contamination detection kit to detect and maintain best practices.
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