The Invitrogen PureLink RNA Mini Kit provides rapid column-based purification of total RNA from a broad range of cell and tissue types. Compatible cell and tissue types include mammalian cells, plant cells, yeast cells, bacteria, and blood.
RNA purified from tissue culture cells using the PureLink RNA Mini Kit is ready for RT-PCR and subsequent downstream analysis by qPCR using gold-standard Applied Biosystems TaqMan gene expression assays. An example qPCR workflow for gene expression analysis using RNA isolated using the PureLink RNA Mini Kit is shown in Figure 1.
Figure 1. An example of PureLink RNA Mini Kit to qPCR workflow. This workflow begins with RNA isolation from tissue culture using the PureLink RNA Mini Kit. The total RNA obtained from the sample is then ready for RT-PCR with the ProFlex PCR System. Relative amounts of RNA between samples are measured using TaqMan assays via qPCR using the QuantStudio 7 Pro Real-Time PCR System.
Depending on the cell or sample type and starting amount, the PureLink RNA Mini Kit extracts an average yield of 1.8–350 µg per sample (Table 1). View the list of cells/sample types compatible with the PureLink RNA Mini Kit in Table 2.
|Sample type||Sample||Starting amount||Average yield (µg)|
|Animal cells||HeLa cells||1 x 106 cells||15–20|
|293 cells||1 x 106 cells||20–25|
|Rat liver||10 mg|
|Rat brain||10 mg|
|Rat spleen||10 mg|
|Calf thymus||10 mg|
|Plant tissue||Arabidopsis||100 mg||26|
|Sugar beet||100 mg||31|
|1 x 107||1.8|
|5 x 108||125|
|Animal and plant cells||Up to 5 × 107 cells|
|Animal tissue||Up to 200 mg|
|Bacterial cells||Up to 109 cells|
|Plant tissue||Up to 250 mg|
|Whole blood||Up to 0.2 mL|
|Yeast cells||Up to 5 x 108 cells|
Column kits are available for total RNA, mRNA, miRNA, small RNA, viral RNA, and sequence-specific RNA extractions. Total RNA isolated using the PureLink RNA Mini Kit can be used in many downstream applications, which require high-quality total RNA (Figure 2). These downstream applications include:
Figure 2. The look of high-quality RNA. RNA was purified from 1 x 106 HeLa cells TRIzol Reagent, the ChargeSwitch Total RNA Cell Kit, the PureLink RNA Mini Kit, and the TRIzol Plus Purification System. (A) A260/A280 and A260/A230 and (B) RNA Integrity Numbers (RIN) were assessed using a spectrophotometer and an Agilent Bioanalyzer.
If removal of residual DNA is necessary, the PureLink RNA Mini Kit offers on-column treatment to save time and precious RNA samples, or post-isolation DNase treatment with the PureLink DNase Set.
PureLink technology combines non-toxic lysis with the speed, purity and ease of use of silica-membrane column RNA purification. The safe and easy RNA purification procedure can be completed in less than 20 minutes, and high-quality total RNA can be obtained from mini- to midi-prep amounts of starting material with no genomic DNA contamination.
Figure 3. Silica-based purification protocol using PureLink RNA Mini Kit in conjunction with TRIzol Reagent.
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