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Kits
Meaningful
RNA Results
Co-Extraction
of mRNA and DNA
To order or get further information, contact ASI at
865-483-1113 or email us.
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Purified
Nucleic Acid Recovered from a Forest A-Horizon Soil
Sample. Look closely! The TruRNA
product is so clean, it is nearly invisible.
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Suitable
for all samples, even those from carbon-rich environments
The perfect tool for
extracting nucleic acids from any type of soil
Unique purification
process produces RNA that properly responds to enzymatic manipulation
Low-temperature
lysing provides mRNA that is representative of the environment at the
time of sampling
Long nucleic acid product
(100 kb) is perfect for cloning
DNA and RNA are
extracted together
Available
in two sizes: the Mini-Kit
(25 preparations for samples up to 250 mg each) or the Mid-Scale Kit
(15 preparations for samples up to 2.5 g samples with high total organic carbon
content or
up to 5 g samples with low total organic content) |
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The TruRNA
kit produces high-purity nucleic acids with all discoloration
removed as shown in the picture above. Nuclease analysis shows that
the discolored component contains DNA (see next bullet).
The
Figure at right shows electrophoretic profiles of unpurified and TruRNA-purified total nucleic acids
extracted from two deciduous forest A-horizon soil samples (1-2),
two flood plain sediment samples (3-4), and two grassy rhizosphere
soil samples (5-6). The component removed by TruRNA
purification
contains fragmented DNA, suggesting that it is likely extracellular DNA at
some stage of the humification process. It is a potent PCR
inhibitor.
Note the excellent stoichiometry of the 23S and
16S ribosomal RNA bands.
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TruRNA
Enables Efficient
Enzymatic Manipulation!
Every Sample! Every Time!
üThe
electrophoresis gel images at right compare glnA RTPCR products generated using
TruRNA purification and purification from a competitor's kit on
deciduous forest A-Horizon soil samples.
TruRNA
kits were
used to extract the nucleic acid. Final purification was
performed by the TruRNA
kit and by a competitor's kit. RNA was
separated from the DNA, quantified, and a two-fold dilution
series was prepared. Reverse transcription was then performed on
each of the dilutions.
After PCR, gel
electrophoresis images were obtained (right). The lanes correspond
to approximately the following amounts of RNA in the reverse
transcription reaction:
(1) 1000 ng
(2) 500 ng
(3) 250 ng
(4) 125 ng
(5) 63 ng
(6) 1000 ng RT negative control
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RTPCR Comparison of Purification Methods
TruRNA & Competitor
TruRNA
Deciduous Forest
A-Horizon
Soil
M 1
2
3 4
5 RT-
Competitor
M 1
2
3 4
5 RT-
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The TruRNA extraction system has been designed
to extract highly purified RNA and DNA from a wide variety of samples, including all types of soils. Extensive research
has yielded a system that is ideal for preparation of total nucleic
acids including functional gene messenger RNA (mRNA) from even the most carbon-rich environments.
TruRNA
recovers mRNA that accurately represents the sample at the time
it was acquired and frozen. DNA is recovered simultaneously. Until the advent of
the TruRNA kit, it has been particularly difficult to evaluate the expressed
gene content in many samples (e.g. environmental samples) because typical
half-lives of prokaryotic mRNA are measured in seconds to minutes. For this
reason, samples must be frozen in liquid N2 immediately upon
acquisition and maintained in cryo-storage until the nucleic acids are
extracted. However, conventional extraction methods require the sample to be
brought into solution prior to cell disruption, a process which allows a change
in the expressed mRNA levels in the sample and makes meaningful interpretation
of the results extremely difficult. The TruRNA system is designed
so that samples are not thawed or brought into suspension prior to inactivation
of the cellular machinery.
Humic acids and fulvic acids are easy to separate from
DNA but this is not true of environmental RNA, particularly soil RNA. The unique
purification system used in the TruRNA system produces microbial
community RNA that is amenable to reverse transcription and other
molecular manipulations. The TruRNA extraction system has been proven to
effectively recover nucleic acids from even the toughest microbial types
including fungi, actinomycetes, and small Gram-positive cocci including Rhodococcus
and Micrococcus species.
For
many areas of molecular ecology research, such as microbial community dynamics with determination of the expressed
microbial community gene complement, the TruRNA
system has no match. It
is the only reagent system that recovers environmental mRNA that is truly
representative of what was expressed by the microbial community at the time of
environmental sample acquisition. Methods that require suspension of
environmental samples prior to environmental soil RNA extraction allow a change
in the expressed levels of microbial community mRNA. The TruRNA total nucleic
acid extraction system offers lysis and elimination of all enzyme activity below
temperatures where enzymatic processes occur.
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TruRNA
Kits - Capacity, availability and pricing
MiniKit
- 25 preparations (suitable for up to 250 mg samples) - $250
Mid-Scale Kit - 15 preparations (suitable for up to 2.5 g samples with high total organic carbon content;
up to 5 g samples with low total organic content) - $250.
Kits are usually shipped in one to three working days. Contact
us for special shipping requirements.
TruRNA
Downloads
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Requirements
for TruRNA Kits
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To
use the TruRNA Kits, the user should have available
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Mortar / Pestle (appropriate for size of sample) |
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Liquid
N2 |
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Pipettors
(20-, 100-, & 1000-ml
sizes), Balance, Spatula, Gloves |
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Centrifuge (minimum range of 500 to
16,000 ´
g); microcentrifuge is acceptable for the Minikit, the Mid-Scale Kit requires a high-speed
centrifuge. |
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50 ml polypropylene centrifuge tubes
(Mid-Scale Kit only). |
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50 ml Teflon
Oak Ridge tubes (Mid-Scale Kit only). |
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2-mercaptoethanol (approximate 100 ml
per kit) |
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Chloroform:isoamyl alcohol 49:1 (v/v;
reagent grade) – 2 ml per sample.
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Isopropyl alcohol (reagent grade) –
approximately 20 ml per sample. |
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Reagent system for fractionation of
total nucleic acids (if fractionation is desired).
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Look for a combined
extraction/purification and RNA/DNA fractionation system in
upcoming versions of
the TruRNA environmental molecular analysis package. |
To order or get further information, contact ASI at
865-483-1113 or email us.
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(865) 483-1113 (voice), (865) 483-3316 (FAX)