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.

RELATED PRODUCTS:

• RNase A (Cat# M1227-25)
• RNaseOFF ribonuclease Inhibitor (Cat# M1238-4000)
• DNase, E.coli DNA Ligase (Cat# M1217-100)
• Hyperlink-FAST 5 Minutes DNA Ligation Package deal (Cat# Okay902-50)
• New T4 DNA Ligase (Cat# M1247-200)
• T4 DNA Ligase (5 u/μl) (Cat# 9101-250)
• T4 RNA Ligase 1 (ssRNA Ligase) (Cat# M1218-100)
• T4 RNA Ligase 2 (dsRNA Ligase) (Cat# M1219-100)

RNase H (RNase-free)
K1093-5000	ApexBio	5000U	EUR 150
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 A
abx098139-1ml	Abbexa	1 ml	EUR 260.4

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

PI/RNAse
PI/RNASE	ImmunoStep	200 test	EUR 208.2

RNase A
M1227-25	Biovision	each	EUR 294

RNase A
RN103-001	GenDepot	300mg	EUR 3138

RNase A
RB0473	Bio Basic	500mg	EUR 219.73

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

RNase H
RK20535	Abclonal	250U	EUR 88

RNase FREE, 250ml Bottle Removes RNase & DNAse
10-172	Genesee Scientific	1 Bottle/Unit	EUR 269

Rnase (90–105)
5-01885	CHI Scientific	4 x 1mg	Ask for price

Rnase A, 250mg
R9001-250mg	ACTGene	each	EUR 337.2

RNase Inhibitor
EQ010	ELK Biotech	1000U	EUR 33.5

RNase FREE, 1 Liter Bottle Removes RNase & DNAse
10-173	Genesee Scientific	1 Bottle/Unit	EUR 337

RNase FREE, 4 Liter Bottle Removes RNase & DNAse
10-456	Genesee Scientific	1 Bottle/Unit	EUR 403

RNase FREE, 475ml Spray Bottle Removes RNase & DNAse
10-228	Genesee Scientific	1 Spray Bottle/Unit	EUR 300

Rnase A, 1g
R9001-1g	ACTGene	each	EUR 986.4

Rnase A, 1g
R9001	ACTGene	1g	EUR 667.9

RNase A (MBG)
409-050	GeneOn	50 mg	EUR 87.6

RNase A (MBG)
409-250	GeneOn	250 mg	EUR 231.6

RNase L Antibody / Rnasel
RQ6435	NSJ Bioreagents	100 ug	EUR 356.15
Description: RNase L is an enzyme that in humans is encoded by the RNASEL gene. This gene encodes a component of the interferon-regulated 2-5A system that functions in the antiviral and antiproliferative roles of interferons. Mutations in this gene have been associated with predisposition to prostate cancer and this gene is a candidate for the hereditary prostate cancer 1 (HPC1) allele.

RNase III, E.coli
M1226-100	Biovision	each	EUR 268.8

RNase L Peptide
46-735P	ProSci	0.1 mg	EUR 405.6
Description: ABCE1 / RNase L Peptide

RNase T2 Antibody
abx237324-100ug	Abbexa	100 ug	EUR 610.8

RNase 1 Antibody
20-abx141499	Abbexa	EUR 444.00 EUR 727.20 EUR 360.00	100 ul 200 ul 50 ul

RNase 13 Antibody
20-abx141582	Abbexa	EUR 444.00 EUR 727.20 EUR 360.00	100 ul 200 ul 50 ul

RNAse H2A Peptide
4979P	ProSci	0.05 mg	EUR 197.7
Description: (IN) RNAse H2A peptide

RNase A Solution
G118	ABM	1.0 ml; 10 mg/ml	EUR 122.4