EXTRAClean Urine Cell-Free Circulating RNA Purification Kits

Supporting Data

Figure 1. Average read quality distribution relative to raw reads of small RNA sequencing of cell-free RNA urine sample using Urine Cell-Free Circulating RNA Purification kits. Calculations were relative to raw reads.

Figure 2. Genome mapping distribution relative to input reads obtained from small RNA sequencing of cell-free RNA urine sample using Urine Cell-Free Circulating RNA Purification kits.

Figure 3. Average small RNA biotype distribution relative to the input reads obtained from sequencing of cell-free RNA urine sample using Urine Cell-Free Circulating RNA Purification kits.

Kit Specifications
Sample Type	Fresh or frozen urine
Sample Volume Range	250 μL to 2mL
Minimum Elution Volume	50 μL
Maximum Elution Volume	100 μL
Time to Complete 10 Purifications	25–30 minutes
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Average Yields ¥	Variable depending on specimen

^¥Please check page 5 of protocol for average urine yields and common RNA quantification methods.

Storage Conditions and Product Stability

All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Component	Mini (Cat. 72900)	Midi (Cat. 72910)	Maxi (Cat. 72920)
Binding Solution K	25 mL	75 mL	1 x 75 mL 1 x 25 mL
Lysis Buffer A	30 mL	20 mL	20 mL
Wash Solution A	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
EXTRAClean Mini Spin Columns	50	20	10
EXTRAClean Midi Spin Columns	-	20	-
EXTRAClean Maxi Spin Columns	-	-	10
Product Insert	1	1	1

Documentation

PROTOCOLS

(73900) EXTRAClean Urine Cell-Free Circulating RNA Purification Kit (Mini) - Protocol (50 preps)

(73910) EXTRAClean Urine Cell-Free Circulating RNA Purification Kit (Midi) - Protocol (20 preps)

(73920) EXTRAClean Urine Cell-Free Circulating RNA Purification Kit (Maxi) - Protocol (10 preps)

FAQs

Mini, Midi, Maxi

What if a variable speed centrifuge is not available?

A fixed speed centrifuge can be used, however reduced yields may be observed.

At what temperature should I centrifuge my samples?

All centrifugation steps are performed at room temperature. Centrifugation at 4°C will not adversely affect kit performance.

What if I added more or less of the specified reagents’ volume?

Adding more or less than the specified volumes may reduce both the quality and the quantity of the purified RNA. Eluting your RNA in high volumes will increase the yield but will lower the concentration. Eluting in small volumes will increase the concentration but will lower the overall yield.

What if I forgot to do a dry spin before my final elution step?

Your purified RNA will be contaminated with the Wash Solution A. This may reduce the quality of your purified RNA and will interfere with your downstream applications.

Can I perform a second elution?

Yes, but it is recommended that the 2nd elution be in a smaller volume (50% of 1st Elution). It is also recommended to perform the 2nd elution into a separate elution tube to avoid diluting the 1st elution.

Why do my samples show low RNA yield?

Urine samples contain very little RNA. This varies from individual to individual based on numerous variables. In order to increase the yield, the amount of urine input could be increased.

Why do the A260:280 ratio of the purified RNA is lower than 2.0?

Most of the Free-Circulating Urine RNA is short RNA fragments. The A260:280 ratio is normally between 1—1.6. This low A260:280 ratio will not affect any downstream application.

Why does my isolated RNA not perform well in downstream applications?

If a different Elution Buffer was used other than the one provided in the kit, the buffer should be checked for any components that may interfere with the application. Common components that are known to interfere are high salts (including EDTA), detergents and other denaturants. Check the compatibility of your elution buffer with the intended use.

EXTRAClean Urine Cell-Free Circulating RNA Purification Kits