For the purification of total RNA - including microRNA - with rapid gDNA removal
Norgen's Total RNA Purification Plus Kit provides a rapid method for the isolation and purification of total RNA in as little as 20 minutes. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), without the use of phenol or chloroform. Genomic DNA is removed from the sample using a Genomic DNA Removal column, and the RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform. Norgen's kit is the only kit on the market that isolates true total RNA, as other kits must use phenol to recover all sizes of RNA.
Norgen's Total RNA Isolation Plus Kit is unique in that it contains a Genomic DNA Removal Column, allowing for gDNA removal without the use of enzymes. HeLa cells were either passed through the gDNA Removal Column, or received no gDNA removal treatment. The Genomic DNA Removal Column successfully removed gDNA from both 1 million HeLa cells (Panel A) and 100,000 cells (Panel B). For both Panels A and B, the left image displays the RNA run on a 1.2% EtBr-agarose gel, to better visualize any gDNA contamination. The right image displays a 1.0% formaldehyde-agarose gel to allow for better denaturation of RNA in the sample. For higher inputs, as in image A, the Genomic DNA Removal Column also resulted in higher RNA yields than the non-treated samples.
Genomic DNA is well known to contaminate RNA samples, and subsequently RT-qPCR results. Therefore, to determine the amount of genomic DNA contamination in RNA samples, an RT-qPCR can be used to compare Ct values observed with and without reverse transcription of the RNA sample. This is known as the Delta (Δ) Ct. One million and 100,000 HeLa cells were either passed through the gDNA Removal Column or subjected to DNase-treatment using Norgen's RNase-Free DNase I (Cat. No.: 25710). The gDNA Removal column eliminated gDNA similar to a DNase treatment. For higher inputs (i.e. 1 Million HeLa cells), the gDNA Removal Column successfully removed more gDNA contamination from the RNA samples than the DNase-treatment. When inputs were lowered, the amount of genomic DNA contamination in the DNase-treated sample was similar to the sample passed through the gDNA Removal Column. Both methods work well for removing gDNA contamination in RNA samples.
|Maximum Binding Capacity||50 µg|
|Maximum Loading Volume Per Spin Column||650 µL|
|Size of RNA Purified||All sizes, including < 200 nt|
|Time to Complete 10 Purifications||20 minutes|
|RNA Yield||HeLa (1 x 106): 15 µg
E coli (1 x 109): 50 µg