Description
The Monarch Total RNA Miniprep Kit is a comprehensive solution for sample preservation, cell lysis, gDNA removal, and purification of total RNA from a wide variety of biological samples, including cultured cells, blood, and mammalian tissues. Additionally, tough-to-lyse samples, such as bacteria, yeast, and plant, can be processed with additional steps that enhance lysis. Cleanup of enzymatic reactions or purification of RNA from TRIzol® -extracted samples is also possible using this kit. Purified RNA has high quality metrics, including A260/280 and A260/230 ratios >1.8, high RIN scores, and minimal residual gDNA. Captured RNA ranges in size from full-length rRNAs down to intact miRNAs. Additionally, differential binding conditions allow selective capture or exclusion of the sub-200 nucleotide RNA pool that includes miRNA, 5S rRNA, and tRNA. Purified RNA is suitable for downstream applications, such as RT-qPCR, cDNA synthesis, RNA-seq, Northern blot analysis, etc.
Workflow for the Monarch Total RNA Miniprep Kit
Properties:
| Binding Capacity: |
100 µg RNA |
| RNA Size: |
≥ 20 nt |
| Purity: |
A260/280 typically ≥ 2.0, A260/230 usually ≥ 1.8 |
| Tissue Input: |
up to 20 mg for high yield tissues |
| |
up to 50 mg for low yield tissues |
| Cultured Mammalian Cells and Blood Cells (PBMC's, WBC's) Input: |
up to 1 x 106 |
| Mammalian Whole Blood Input: |
up to 3 ml |
| Bacteria Input: |
up to 1 x 109 |
| Yeast Input: |
up to 5 x 107 |
| Plant Input: |
up to 100 mg |
| Elution Volume: |
30 - 100 μl |
| Yield: |
varies depending on sample type |
| Compatible downstream applications: |
RNA Library prep for NGS, RT-PCR, RT-qPCR, Northern blots |
Tested Sample Types:
- HeLa Cells
- HEK 293 Cells
- NIH 3T3 Cells
- Human Blood
- Rat Blood
- PBMC's
- Plasma
- Serum
- Nucleated Blood
|
- Rat Liver
- Rat Spleen
- Rat Kidney
- Rat Brain
- Rat Muscle
- Mouse Muscle
- Mouse Heart
- Mouse Kidney
- Zebrafish Larvae
|
- D. melanogaster
- S. cerevisiae
- E. coli
- B. cereus
- Tomato Leaf
- Corn Leaf
- Saliva
- Buccal cells
|
Additional Data Figures:
Figure 1. Monarch-purified RNA is high-quality and compatible with a wide variety of downstream applications
Total RNA from a broad array of sample types was purified using the Monarch Total RNA Miniprep Kit (NEB #T2010). Aliquots were run on an Agilent® Bioanalyzer® 2100 using the Nano or Pico 6000 RNA chip (S. cerevisiae RNA was run using a plant Nano assay). RIN values and O.D. ratios confirm the overall integrity and purity of the RNA. To demonstrate compatibility with downstream applications, samples were subsequently used for RT-PCR (A) using Protoscript® II Reverse Transcriptase (NEB #M0368)/LongAmp® Taq DNA Polymerase (NEB #M0323), NGS library prep (B) using NEBNext® Ultra™ II RNA Library Prep Kit (NEB #E7760) and RT-qPCR (C) using Luna qPCR Reagents (NEB #E3005).
Figure 2. The Monarch Total RNA Miniprep Kit can generate high quality RNA from as few as 100 HeLa cells
Total RNA was isolated using the Monarch Total RNA Miniprep Kit (NEB #T2010) from varying amounts of HeLa cells over 5 orders of magnitude and eluted in 100 µl of nuclease-free water. Samples were analyzed on a Bioanalyzer Pico chip, with RIN values and total yields shown below the lanes (A). Electropherograms are included as a reference (B).
Figure 3. The Monarch Total RNA Miniprep Kit successfully purifies small RNAs below 200 nucleotides, enabling a more faithful representation of the total RNA pool
RNA preps were performed on HEK293 cells, mouse heart, rat brain, or rat spleen using the Monarch Total RNA Miniprep Kit (NEB #T2010) and the RNeasy Mini Kit from Qiagen. Equivalent amounts were resolved on a Bioanalyzer 2100 using the Small RNA chip. Monarch-purified RNA contains significantly more RNA in the sub-200 nucleotide pool.
- This product is related to the following categories:
-
Discontinued (<3 years),