Choosing Sample Input Types for RNA Purification and Average Yields Expected
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® Spin RNA Isolation Kit (Mini) (NEB #T2110). It is important not to overload the column when extracting and purifying RNA, as yields, purity and integrity may suffer.
|
SAMPLE TYPE |
RECOMMENDED INPUT AMOUNT |
TYPICAL YIELD (µg) |
OBSERVED RIN or RINe |
MAXIMUM INPUT AMOUNT |
|
|---|---|---|---|---|---|
|
CULTURED CELLS |
|||||
|
HEK 293 |
1 x 106 cells |
12–14 |
9–10 |
5 x 106 cells |
|
|
NIH3T3 |
1 x 106 cells |
8–12 |
9–10 |
1 x 107 cells |
|
|
TISSUE |
|||||
|
Rat liver (frozen stabilized) |
10 mg |
40–60 |
8–9 |
20 mg |
|
|
Rat spleen (frozen stabilized) |
10 mg |
40–50 |
9 |
20 mg |
|
|
Rat kidney (frozen stabilized) |
10 mg |
7–10 |
9 |
50 mg |
|
|
Rat brain (frozen stabilized) |
10 mg |
5–8 |
8–9 |
50 mg |
|
|
Rat muscle (frozen stabilized) |
10 mg |
2–3 |
8–9 |
50 mg |
|
|
Mouse heart (frozen stabilized) |
10 mg |
5–6 |
8–9 |
50 mg |
|
|
BLOOD OR PLASMA |
|||||
|
Human |
Fresh |
200 µl |
0.5–1.0 |
7–8 |
200 µl |
|
Frozen |
200 µl |
0.5–1.0 |
7–8 |
200 µl |
|
|
Rat |
Frozen |
200 µl |
5-6 |
9 |
200 µl |
|
BLOOD CELLS |
|||||
|
PBMC (isolated from 5 ml whole blood) |
5 ml |
1-3 |
7 |
5 x 106 cells |
|
|
YEAST |
|||||
|
S. cerevisiae |
Frozen with bead homogenizer |
1 x 107 cells |
20-40 |
9–10 |
5 x 107 cells |
|
Frozen with Zymolyase® |
1 x 107 cells |
20-40 |
9 |
5 x 107 cells |
|
|
BACTERIA |
|||||
|
E. coli |
Frozen pellet with bead homogenizer |
1 x 109 cells |
10-15 |
10 |
1 x 109 cells |
|
Frozen pellet with lysozyme |
1 x 109 cells |
40-60 |
10 |
1 x 109 cells |
|
|
B. subtilits |
Frozen pellet with bead homogenizer |
1 x 109 cells |
15–20 |
9 |
1 x 109 cells |
|
Frozen pellet with lysozyme |
1 x 109 cells |
20-30 |
9–10 |
1 x 109 cells |
|
|
PLANT |
|||||
|
Corn leaf (frozen pulverized with bead homogenizer) |
100 mg |
40-60 |
8* |
100 mg |
|
|
Tomato leaf (frozen pulverized with bead homogenizer) |
100 mg |
40-60 |
8* |
100 mg |
|
|
Onion leaf (fresh with bead homogenizer) |
50 mg |
4-6 |
8* |
50 mg |
|
|
Root |
50 mg |
8-10 |
8* |
50 mg |
|
|
INSECTS |
|||||
|
Mosquito (preserved in ethanol dry ice bath) |
10 mg |
20-30 |
9* |
20 mg |
|
|
House fly (preserved in ethanol dry ice bath) |
10 mg |
10-20 |
9* |
10 mg |
|
RNA yields and RINs observed during evaluation are reported here. RNA yield and quality are influenced by several factors including sample growth stage and RNA content, storage condition, sample handling during processing as well as the chosen method of sample lysis and homogenization. It is crucial to consider these factors and use best practices to maximize RNA yield and quality. Please refer to Important Notes in the Product Manual before starting for recommendations on Working with RNA and Considerations for Sample Lysis and Homogenization.
*These samples contain RNA with an atypical ribosomal profile that is not applicable for standard RIN measurement. Users are encouraged to set their own threshold or assess RNA integrity by visually inspecting the bands generated by the automated gel electrophoresis platform.
