Guidance on Choosing Sample Input Amounts when Using the
Monarch Total RNA Miniprep Kit
RNA yield, purity, and integrity vary immensely based on sample type, input amount and sample condition. Below, we have provided some empirical yield, purity, and RIN data from a wide variety of sample types, as well as guidance on the maximal input amounts for each of those samples when using the Monarch Total RNA Miniprep Kit (NEB #T2010). It is very important not to overload the column when extracting and purifying RNA, as yields, purity and integrity may suffer. If you have any specific questions regarding sample inputs, please don’t hesitate to contact our technical support staff.
| Sample Type† | Input | Average Yield (μg) | Observed RIN | Maximum Starting Material | |
|---|---|---|---|---|---|
| Cultured mammalian cells | |||||
| HeLa | 1 x 106 cells | 12–15 | 9–10 | 1 x 106 cells | |
| HEK 293 | 1 x 106 cells | 12–14 | 9–10 | 1 x 106 cells | |
| NIH 3T3 | 1 x 106 cells | 8–12 | 9–10 | 1 x 106cells | |
| Mammalian blood† † | |||||
| Human | Fresh | 200 μl | 0.5–1.0 | 7–8 | 3 ml |
| Frozen | 200 μl | 0.5–1.0 | 7–8 | 3 ml | |
| Stabilized | 200 μl | 0.5–1.0 | 7–8 | 3 ml | |
| Rat | Frozen | 100 μl | 5.6 | 9 | 1 ml* |
| Blood cells | |||||
| PBMC (isolated from 5 ml whole blood) | 5 ml | 3 | 7 | 1x106 cells | |
| Tissue | |||||
| Rat liver | Frozen pulverized | 10 mg | 25 | 8–9 | 20 mg |
| Stabilized solid | 10 mg | 50–60 | 8–9 | 20 mg | |
| Rat spleen (stabilized solid with bead homogenizer) | 10 mg | 40–50 | 9 | 20 mg | |
| Rat kidney (Frozen pulverized) | 10 mg | 7–10 | 9 | 50 mg | |
| Rat brain | Frozen pulverized | 10 mg | 2–3 | 8–9 | 50 mg |
| Stabilized solid | 10 mg | 0.5–1.5 | 8–9 | 50 mg | |
| Stabilized solid with bead homogenizer | 10 mg | 5–8 | 8–9 | 50 mg | |
| Rat muscle (Frozen pulverized) | 10 mg | 2–3 | 8–9 | 50 mg | |
| Mouse muscle | Frozen pulverized | 10 mg | 3 | 8–9 | 50 mg |
| Frozen pulverized with bead homogenizer | 10 mg | 5 | 7–8 | 50 mg | |
| Stabilized solid with bead homogenizer | 10 mg | 8–10 | 9 | 50 mg | |
| Mouse heart (stabilized solid w/ bead homogenizer) | 10 mg | 5–6 | 8–9 | 50 mg | |
| Yeast | |||||
| S. cerevisiae | Frozen with bead homogenizer | 1 x 107 cells | 50 | 9–10** | 5 x 107 cells |
| Fresh with Zymolyase® | 1 x 107 cells | 60 | 9** | 5 x 107 cells | |
| Bacteria | |||||
| E. coli | Frozen | 1 x 109 cells | 5 | 10 | 1 x 109 cells |
| Frozen with bead homogenizer | 1 x 109 cells | 10 | 10 | 1 x 109 cells | |
| Frozen with lysozyme | 1 x 109 cells | 70 | 10 | 1 x 109 cells | |
| B. cereus | Frozen with lysozyme | 1 x 108 cells | 20–30 | 9 | 1 x 109 cells |
| Frozen with bead homogenizer | 1 x 108 cells | 8 | 9–10 | 1 x 109 cells | |
| Plant | |||||
| Corn leaf (frozen pulverized with bead homogenizer) | 100 mg | 45 | 8 | 100 mg | |
| Tomato leaf (frozen pulverized with bead homogenizer) | 100 mg | 30 | 8 | 100 mg | |
† RNA for other samples including drosophila, zebrafish embryos/larvae, plasma, serum, saliva, buccal swabs and nucleated blood have been successfully purified with this kit; protocols are available in the product manual.
†† A protocol for nucleated blood (e.g., birds, reptiles) is also available.
* Mouse blood also has a maximum input of 1ml.
** S. cerevisiae total RNA was run on an Agilent® Nano 6000 Chip using plant assay.
