GDP420-RNAprep Pure Micro Kit

RNAprep Pure Micro Kit
For purification of total RNA from micro samples

RNAprep Pure Micro Kit
(Spin Column)
Cat. no. GDP420

Storage
RNAprep Pure Micro Kit can be delivered at room temperature (15-30°C).
When the kit has arrived, store RNase-free DNase I, Buffer RDD, and
RNase-free ddH2O at 2-8°C for up to 15 months. Other reagents can be
stored at room temperature (15-30°C) for 15 months.

Introduction
RNAprep Pure Micro Kit adopts innovative technology to guarantee the
isolation of RNA from micro amount of tissue and cell samples(as little
as 10 cells). Unique component added in buffer system greatly enhances
the capability of selective binding of RNA to silica membrane. RNase-free
DNase I has been added to remove the trace DNA contamination. DNase
I and other residual impurity will be removed in the following washing
step. Eventually efficient purification of high-quality RNA in a small elution
volume is guaranteed.
With the RNAprep Micro Pure Kit procedure, all RNA molecules longer than
200 bp are purified. The procedure enriches for mRNA, since most RNAs
<200 bp such as 5.8S rRNA, 5S rRNA, and tRNAs are selectively excluded.
In the handbook, different protocols are provided for different
starting materials. The protocols differ primarily in the disruption and
homogenization of the sample. Once the sample is applied to RNase-free
Spin Column CR1, the subsequent steps are similar.

Important Notes

1. Regents and consumables (not supplied):β-mercaptoethanol, ethanol
   (96%-100%), 1.5 ml RNase-free microcentrifuge tube.
2. Determining the amount of starting materials:
   It is essential to use the correct amount of starting material in order to
   obtain optimal RNA yield and purity. The maximum amount that can be
   used is determined by the type of sample and its RNA content. When
   processing samples containing high amounts of DNA or RNA, less than
   the maximum amount of starting material shown in Table 1 should be
   used, Table 2 shows typical RNA yields from various cells.

3. Storage and treatment of samples
   In untreated tissues, RNA is easy to degrade in unprotected environment,
   so fresh tissues should be placed into liquid nitrogen and stored at -70°C
   immediately. Do not freeze-thaw the tissues. Buffer RL can be added to
   tissue samples after homogenization, and then stored at -70°C for several
   months.
4. Disruption and homogenization of samples
   Efficient disruption and homogenization of the starting material is an
   absolute requirement for all total RNA purification procedures. Disruption
   and homogenization are two distinct steps.
   Disruption: Complete disruption of plasma membranes of cells and
   organelles is absolutely required to release all the RNA contained in the
   sample. Different samples require different methods to achieve complete
   disruption. Incomplete disruption results in significantly reduced RNA
   yields. Homogenization is necessary to reduce the viscosity of the lysates
   produced by disruption.
   Homogenization: Shears the high molecular-weight genomic DNA
   and other high-molecular-weight cellular components to create a
   homogeneous lysate. Incomplete homogenization results in inefficient
   binding of RNA to the RNase-free Spin Column CR1 membrane and
   therefore significantly reduces RNA yields. Homogenization could be
   operated by homogenizer, vortex, filtered column, syringe and pipette.
5. Carrier RNA
   The RNAprep Pure Micro Kit contains poly-A RNA for use as Carrier
   RNA. When added to lysates of very small samples, the Carrier RNA may
   in some cases improve the recovery of total RNA. Carrier RNA is not
   required when processing more than 5,000 cells or 10 μg tissue.

GDP420-RNAprep Pure Micro Kit

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This product is for scientific research use only. Do not use in medicine, clinical treatment, food or cosmetics.

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