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 1x10cells 12–15 9–10 1 x 107 cells
HEK 293 1x10cells 12–14 9–10 1 x 107 cells
NIH 3T3 1x10cells 8–12 9–10 1 x 107 cells
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 1x107 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 1x107 cells 50 9–10** 5x107 cells
Fresh with Zymolyase® 1x107 cells 60 9** 5x107 cells
Bacteria
E. coli Frozen 1x109 cells 5 10 1x109 cells
Frozen with bead homogenizer 1x109 cells 10 10 1x109 cells
Frozen with lysozyme 1x109 cells 70 10 1x109 cells
B. cereus Frozen with lysozyme 1x108 cells 20–30 9 1x109 cells
Frozen with bead homogenizer 1x108 cells 8 9–10 1x109 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.