Spectrum™ Plant Total RNA Kit

The total RNA purification method for difficult plant tissues

Few methods are capable of delivering desired amounts of total RNA from plant tissues that contain high levels of secondary metabolites. The Spectrum™ Plant Total RNA Kit was designed to be different. Incorporating a new, patent-pending lysis and binding chemistry, this kit is intended to purify total RNA from species and tissue where most protocols fail. Without sacrificing affordability, you can purify more total RNA from difficult tissue samples.

The Spectrum™ Plant Total RNA Kit employs a new lysis and binding chemistry and a convenient column-based 'bind-wash-and-elute' format to purify up to 100 µg of total RNA from 100 mg of tissue in about 30 minutes. Typical yields range from 20–60 µg. After grinding tissue to a fine powder in liquid nitrogen, cells are lysed and cellular debris is physically and chemically separated from endogenous RNA. RNA is then bound to a column supported silica substrate and several wash steps remove remaining contaminants. Lastly, total RNA is eluted from the column and used in typical applications, such as Northern Blots, and RT- and qRT-PCR. The Spectrum™ Plant Total RNA Kit contains all the reagents, buffers and solutions necessary to perform the method as described.

Spectrum™ Plant Total RNA

Total RNA Extraction from Sugarcane Leaf using Spectrum™ Total RNA kit

RNA in all samples extracted from 100 mg tissue and eluted in 60 μL elution buffer, 5 μL loaded on 1.5% denaturating formaldehyde gel. Lanes (1) RNA Marker; (2) Tender Leaf; (3) Mature Leaf; (4) Leaf Sheath; (5) Stem; (6) Auxiliary Bud-Closed; (7) Auxiliary Shoot-Growing.

Total RNA Extraction from Rice using Spectrum™ Total RNA kit

RNA extracted from 100mg tissue, eluted in 60 μL and 4 μL loaded on (A) 2% denaturating formaldehyde gel & (B) TAE native gel. Lanes (1) RNA Marker R7020; (2) Flower - unopened; (3) Root - lateral; (4) Leaf; (5) RNA Marker R7020.

Total RNA Extraction from Cotton using Spectrum™ Total RNA Kit

RNA extracted from 100mg tissue, eluted in 60 μL and 4 μL loaded on 2% denaturing formaldehyde gel. Lanes (1) Leaf; (2) Root; (3) Petal; (4) Seed; (5) Stigma; (6) RNA Marker R7020.

Total RNA Isolation from different tissues of Brinjal using the Spectrum™ Total RNA kit

100 mg of tissue was used in each case from a 70-day-old plant. This 2% Formaldehyde Gel shows results from 60 µL elution of which 4 µL was loaded. Lane (1) Petal-yield 20.4 µg RNA; (2) Anther-yield 16.8 µg RNA; (3) 7 month old Seed-yield 3.8 µg RNA; (4) Leaf-yield 122 µg RNA; (5) Root (lateral)-yield 27.8 µg RNA; (6) RNA marker R7020.

Total RNA Isolation from different ages of rice leaves using the Spectrum™ Total RNA kit

100 mg of tissue was used in each case from a 100-day-old plant. This 2% Formaldehyde Gel shows results from 60 µL elution of which 4 µL was loaded. Lane (1) 60 day old top most rice leaf-yield 79.2 µg RNA; (2) 60 day old lower most rice leaf-yield 62.9 µg RNA; (3) RNA marker R7020; (4) 110 day old top most rice leaf-yield 48.2 µg RNA; (5) 110 day old lower most rice leaf-yield 12.0 µg RNA.

Total RNA Isolation from different ages of cotton leaves using the Spectrum™ Total RNA kit

100 mg of tissue was used in each case from a 100-day-old plant. This 2% Formaldehyde Gel shows results from 60 µL elution of which 4 µL was loaded. Lane (1) 4 month old top most cotton leaf-yield 94.1 µg RNA; (2) 4 month old lower most cotton leaf-yield 30.2 µg RNA; (3) RNA marker R7020; (4) 6 month old top most cotton leaf-yield 83.7 µg RNA; (5) 6 month old lower most cotton leaf-yield 48.7 µg RNA.

Total RNA Isolation from different tissue types of chilli using the Spectrum™ Total RNA kit

100 mg of tissue was used in each case from a 90-day-old plant. This 2% Formaldehyde Gel shows results from 60 µL elution of which 4 µL was loaded. Lane (1) RNA marker R7020; (2) 1 month old chilli seed (3) Root; (4) Leaf; (5) Bud.

Effect of Storage on RNA quality - Total RNA Isolation from rice and cotton using the Spectrum™ Total RNA kit

Plant RNA was stored at –20 °C for 70 days. This 2% Formaldehyde Gel shows results from 60 µL elution of which 4 µL was loaded. Lane (1) RNA marker R7020; (2) Rice Leaf total RNA; (3) Cotton Leaf total RNA.

Total RNA Isolation from different tissue types of tobacco using the Spectrum™ Total RNA kit

100 mg of tissue was used in this 1.5% Formaldehyde Gel which shows results from 60 µL elution of which 5 µL was loaded. Lane (1) Young Leaf; (2) Root; (3) Seed; (4) Sepal; (5) Opened Stamens; (6) Petal; (7) RNA marker R7020.

Total RNA Isolation from different tissue types of maize using the Spectrum™ Total RNA kit

100 mg of tissue was used in this 1.5% Formaldehyde Gel which shows results from 60 µL elution of which 5 µL was loaded. Lane (1) RNA marker R7020; (2) Young Leaf; (3) Leaf Sheath; (4) Root; (5) Cob Sheath; (6) Silk; (7) Opened Stamens.

Total RNA Isolation from different tissue types of papaya using the Spectrum™ Total RNA kit

100 mg of tissue was used in this 1.5% Formaldehyde Gel which shows results from 60 µL elution of which 5 µL was loaded. Lane (1) RNA marker R7020; (2) Young Leaf; (3) Raw Green Fruit; (4) Stalk; (5) Immature, white Seed.

Total RNA extracted from 100 mg of Jatropha tissue

Elution in 60 µL – of which 4 µL were loaded on a 1.5% formaldehyde gel. Lane (1) RNA marker R7020; (2) Young Leaf; (3) Green Stem; (4) Raw green fruit; (5) Flower.

Additional Information

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