Name: Preserved Blood RNA Purification Kit I (for use with Tempus Blood RNA Tubes)
Brand: Norgen Biotek Corp.
SKU: 43400
Price: 525.000000 USD
Availability: InStock

Supporting Data

Figure 1. Isolation of High Quality Total RNA. Total RNA was isolated in duplicate from 3 mL hamster blood samples collected into Tempus™ RNA Blood Tubes using Norgen’s Preserved Blood RNA Purification Kit I and was subsequently analyzed using an Agilent BioAnalzyer. Panel A shows the electropherogram from the Agilent Bioanalyzer, where the y axis represents fluorescence units and the x axis represents the runtime (s). The bands of the 18S and 28S rRNA fragments are clearly visible and the RNA integrity number score is 9.0. Panel B demonstrates the gel banding pattern of the RNA species purified from hamster blood. Abundant blood transcripts such as the globin mRNAs constitute the additional bands to the two major rRNA bands.

Figure 2. Isolate a Diversity of RNA Species, from large mRNA to microRNA. Total RNA was isolated in duplicate from 3 mL hamster blood samples collected into Tempus™ RNA Blood Tubes using Norgen’s Preserved Blood RNA Purification Kit I and was subsequently used as the template in a RT-qPCR reaction for the detection of various sized RNA. All sizes of RNA were successfully amplified including large RNA (Beta-Actin = green), small RNA, such as the 5S rRNA (blue) and microRNA, miR-21 (red), indicating both the size diversity of the purified RNA, as well as the high quality of the RNA. No template control is shown in yellow.

Figure 3. High Yield of a Diversity of RNA Species. Norgen's Preserved Blood RNA Purification Kit I (for use with Tempus™Blood RNA Tubes) effectively recovers all sizes of RNA from large mRNA to small RNA, including microRNAs. Total RNA was isolated from equal amounts of goat blood collected in Tempus™ Blood RNA Tubes followed by RNA isolation using Norgen's Preserved Blood RNA Purification Kit I and a competitor's kit. The purified RNA was then used as the template in a RT-qPCR for detecting the beta-Actin gene (Left Panel) and for detecting miR-21 (Right Panel). In both graphs the blue lines correspond to Norgen isolated-RNA and the red lines correspond to competitor-isolated RNA. As it can be seen, Norgen's kit isolated higher yields of microRNA, as indicated by the lower Ct values of the blue lines (Right Panel). Also, Norgen's kit successfully isolated similar amount of large RNA to that of the competitor’s kit (Lower Panel) indicating the full diversity of RNA species isolated.

Kit Specifications
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	20 - 30 minutes
Average Yield	5 - 25 μg per 3 mL preserved human blood

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 43400 (50 preps)
Tempus™ Blood RNA Tube Diluent	2 x 90 mL
Lysis Solution	40 mL
Wash Solution	22 mL
Elution Solution	6 mL
Mini Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Documentation

PROTOCOLS

(43400) Preserved Blood RNA Purification Kit I - Protocol (50 prep)

SAFETY DATA SHEETS

43400 - Preserved Blood RNA Purification Kit I (For Use with Tempus Blood RNA Tubes) - SDS

FLYERS

Flyer

RELATED BLOGS

FAQs

Spin Column

Why do I have poor RNA recovery?

Poor RNA recovery could be due to one or more of the following:

Incomplete lysis of blood.
Ensure that the appropriate amount of Lysis Solution was used.
An alternative elution solution was used.
It is recommended that the Elution Solution supplied with this kit be used for maximum RNA recovery.
Ethanol was not added to the lysate.
Ensure that the appropriate amount of ethanol is added to the lysate before binding to the column.
Ethanol was not added to the Wash Solution.
Ensure that 50 mL of 95% ethanol is added to the supplied Wash Solution prior to use.
Insufficient centrifugation time and speed when pelleting the stabilized blood RNA before column purification.
Centrifuge the samples for at least 30 minutes or longer at 4,000 x g or higher. The RNA pellet was lost during decanting Pour off the supernatant slowly and carefully.

I noticed the columns are getting clogged. What causes this?

Column clogging can result from one or combination of the following factors:

Insufficient solubilization of blood.
Ensure that the appropriate amount of Lysis Buffer was used.
Centrifuge temperature is too low.
Ensure that the centrifuge remains at room temperature throughout the procedure. Temperatures below 15°C may cause precipitates to form that can cause the columns to clog.

Why is the RNA degrading?

RNA can be degraded due to following factors:

RNase contamination.
RNases may be introduced during the use of the kit. Ensure proper procedures are followed when working with RNA. Please refer to “Working with RNA” at the beginning of the protocol's user guide.
Procedure not performed quickly enough.
In order to maintain the integrity of the RNA, it is important that the procedure be performed quickly.
Improper storage of the purified RNA.
For short term storage RNA samples may be stored at -20°C for a few days. It is recommended that samples be stored at -70°C for longer term storage.

Why is the RNA not performing well in downstream applications?

If the RNA does not perform well in downstream applications, It may be due to one or more of the following:

RNA was not washed 3 times with the provided Wash Solution A.
Traces of salt from the binding step may remain in the sample if the well is not washed 3 times with Wash Solution A. Salt may interfere with downstream applications, and thus must be washed from the well.
Ethanol carryover.
Ensure that the dry spin under Column Wash in the centrifugation protocol or the extended vacuum in the vacuum protocol is performed in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.

I noticed genomic DNA is contaminated. Why did this happen?

Genomic DNA contamination could occur if large amount of starting material used. Perform RNAse-free DNase I digestion on the RNA sample after elution to remove genomic DNA contamination. It is recommended that Norgen’s RNase-Free DNase I Kit (Cat. 25710) be used for this step.

Preserved Blood RNA Purification Kit I (for use with Tempus Blood RNA Tubes)