Cellexus Limited Cell Culture Biosystems

SAMPLE PROCESSING GUIDELINES AND TROUBLESHOOTING:

• For digestion of full RNA, longer incubations of 2-Three hours are generally required.
• If degradation is inefficient, use a barely elevated incubation temperature (40-45°C) and complement additional enzyme partway (e.g. 0.5 μl after 1 hour) by way of the method. The higher temperature is particularly useful for degrading extraordinarily structured linear RNAs, equal to rRNAs. Do not exceed 45°C or incubate over 3
hours, as this may end in non-enzymatic RNA degradation.

• RNase R shows low train on tRNA, rRNA and totally different extraordinarily structured RNAs, for which the three’ end is double stranded with a short 3’ overhang. These RNA species can stall the enzyme and finish in drastically lowered train. If inefficient degradation is seen, it is endorsed to each upscale the digestion, use additional RNase R,
or take away rRNA from full RNA extracts earlier to digestion.

• Keep in mind that spherical RNAs symbolize a small proportion of full RNA (generally 0.1%-0.01%), because of this reality RNase R treatment will most actually finish in low ranges of RNA (picogram-range), presumably undetectable by most methods. For that purpose, a starting amount of not lower than 10 µg of full RNA is useful for a lot of downstream features.

• Whereas the enzyme could also be heat inactivated the method should not be useful since extreme heat can lead to RNA hurt. Phenol-chloroform precipitation could be utilized instead. For NGS, robust half reversible immobilization (SPRI) bead cleanup is useful.

• Magnesium at concentrations of 0.1-1.Zero mM is required for optimum train. If ETA is
present, compensate by together with MgCl2 to 1.Zero Mm final focus.

DESCRIPTION:

RNase R is an E. coli exoribonuclease which shows 3’-to-5’ exonuclease train, successfully digesting virtually all linear RNA species. This enzyme does not digest spherical, lariat, or double stranded RNA with temporary 3’ overhangs (decrease than sevennucleotides). As such, this enzyme is ideally suited to the study of lariat RNA produced by typical splicing, along with circRNAs which come up by way of back-splicing.

APPLICATIONS:
• Enriching circRNAs in natural samples
• Identification of intronic lariat sequences
• Identification of exonic circRNAs
• Studying varied splicing
• Manufacturing of artificial spherical RNAs

EZYME UNIT DEFINITION: One unit is printed as the amount of RNase R that converts 1
µg of poly(A) into acid soluble nucleotides in 10 minutes at 37°C. ENZYME STORAGE BUFFER: 50 mM Tris-HCl (pH 7.5), 100mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 50% (v/v) Glycerol 10X RNase R REACTION BUFFER COMPONENTS: 200 mM Tris-HCl, 1 M KCl, 1 mM, MgCl2, pH 7.5.

STORAGE CONDITIONS: Retailer at -20°C.

Stay away from repeated freeze-thaw cycles of all components to retain most effectivity. All components
are regular for one 12 months from the date of transport when saved and handled appropriately.

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• New T4 DNA Ligase (Cat# M1247-200)
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• T4 RNA Ligase 1 (ssRNA Ligase) (Cat# M1218-100)
• T4 RNA Ligase 2 (dsRNA Ligase) (Cat# M1219-100)

RNase R (20 U/μl)
RNR-E5146	ACROBIOSYSTEMS	2KU	EUR 716.9
Description: RNase R (20 U/μl) is expressed from Escherichia coli K12 E. coli.

RNase H (RNase-free)
K1093-1000	ApexBio	1000U	EUR 42
Description: RNase H

RNase H (RNase-free)
K1093-500	ApexBio	500U	EUR 24
Description: RNase H

RNase H (RNase-free)
K1093-5000	ApexBio	5000U	EUR 120
Description: RNase H

RNAse
E16FXP0211R-200	EnoGene	200 μl	EUR 13.33
Description: Available in various conjugation types.

RNAse
E16FXP0211R-400	EnoGene	400 μl	EUR 20
Description: Available in various conjugation types.

RNAse
E16FXP0211R-800	EnoGene	800 μl	EUR 26.67
Description: Available in various conjugation types.

RNase
TRI-A40S1	TRI Biotech	100UL	EUR 37.8

Rnase (90-105)
MBS8247015-10mg	MyBiosource	10mg	EUR 715

Rnase (90-105)
MBS8247015-1mg	MyBiosource	1mg	EUR 260

Rnase (90-105)
MBS8247015-5mg	MyBiosource	5mg	EUR 510

Rnase (90-105)
MBS8247015-5x10mg	MyBiosource	5x10mg	EUR 3140

RNase A
abx098139-1ml	Abbexa	1 ml	EUR 260.4

PI / RNAse
abx290029-200tests	Abbexa	200 tests	EUR 326.4

RNase A
MB087-100MG	EWC Diagnostics	1 unit	EUR 34.02
Description: RNase A

RNase A
MB087-1G	EWC Diagnostics	1 unit	EUR 284.22
Description: RNase A

RNase A
MB087-500MG	EWC Diagnostics	1 unit	EUR 153.8
Description: RNase A

RNase A
RM7705-100MG	EWC Diagnostics	1 unit	EUR 136.11
Description: RNase A

RNase A
RM7705-1G	EWC Diagnostics	1 unit	EUR 1102.26
Description: RNase A

RNase A
RM7705-500MG	EWC Diagnostics	1 unit	EUR 612.22
Description: RNase A

RNase A
RM1555-100MG	EWC Diagnostics	1 unit	EUR 118.89
Description: RNase A

RNase A
RM1555-500MG	EWC Diagnostics	1 unit	EUR 537.26
Description: RNase A

RNase I
E1300-01	EURx	10000U	EUR 75.21
Description: Completely nonspecific RNase that hydrolyzes phosphodiester bond after all four bases

RNase I
E1300-02	EURx	50000U	EUR 301.93
Description: Completely nonspecific RNase that hydrolyzes phosphodiester bond after all four bases

RNase A
G117	ABM	25 mg	EUR 55
Description: RNase A is pancreatic bovine endoribonuclease that cleaves single stranded RNA by hydrolysis at cytosine and uracil residues. RNase A is very useful for the removal of RNA from plasmid DNA preparations.

RNase A
IB47163	IBI Scientific	50ul	EUR 20.72
Description: (FOR HI-SPEED PLASMID) [FR}

RNase A
M1227-25	Biovision	each	EUR 294

RNase A
RB0473	Bio Basic	500mg	EUR 219.73

RNase H
RK20535	Abclonal	250U	EUR 88

RNase A
RN103-001	GenDepot	300mg	EUR 3138

RNase A
abx098139-100l	Abbexa	100 µl	EUR 250