GUIDELINES

Guidelines and Recommendations
for miRIDIAN™ microRNA Libraries

Handling and storage recommendations
Dharmacon™ miRIDIAN™ reagents are shipped as dry pellets at ambient
temperature and should be stored at –20 °C upon arrival in a manualdefrost or non-cycling freezer. Under these conditions, the mimics andhairpin inhibitors are stable for at least one year. If necessary, dry RNA pellets(unopened) can be stored at 4 °C for several weeks.

Resuspension recommendations
1. Briefly centrifuge plates to ensure that the RNA pellet is collected at thebottom of the well.
2. Wipe adhesive foil cover with 70% ethanol or other RNasedecontamination solution such as Fisherbrand™ RNase Displace™
Decontaminant (Cat #04-355-136; 04-355-138; 04-355-137).
3. Carefully peel back the foil seal to gain access to wells.
Use caution and avoid shredding the seal.
4. Dilute 5x siRNA resuspension buffer (Cat #B-002000-UB-100) to
1x concentration with RNase-free water before resuspending
miRIDIAN products. RNase-free water is available from Dharmacon(Cat #B-003000-WB-100).
Note: For optional RNA quantitation by UV spectrophotometry(at 260 nm), resuspend well(s) in 4 volumes of RNase-free water.
Following this analysis, add 1 volume 5x buffer for appropriate
final 1x concentration. Salts present in buffer are known to cause adecrease in the absorbance reading of RNA. For additional tips on
accurate spectrophotometry readings, please see the FAQ sectionof our website.
5. Resuspend mimics or inhibitors to a convenient stock concentration
using the recommended volume of 1x siRNA buffer or RNase-freewater shown in Table 1. Concentrated stocks of 20 μM or more arerecommended. However, stock solutions of 1-10 μM may betteraccommodate dilution schemes for high-throughput transfections andassays conducted on robotic platforms.

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6. Pipette solution up and down 3-5 times while avoiding introductionof bubbles.
7. Place the solution on an orbital mixer/shaker for 70–90 minutes at room
temperature. This additional mixing results in more reliable resuspension.
8. Briefly centrifuge plates to collect solution to bottom of the wells.
9. Mimics or inhibitors may now be used immediately, stored at –20 °C(4 °C is suitable for 4–6 weeks) in a manual defrost or non-cycling freezer, or
aliquoted into daughter plates.
a. Polypropylene accommodates storage at –80 °C and is often used fordaughter plate creation.
b. Polystyrene plates are suitable for –20 °C storage, but become brittle at-80 °C and may be subject to breakage.
10. Seal plates with appropriate adhesive or heat seal.
11. Limit freeze-thawing of each plate. Up to 15 freeze thaws can be tolerated
but for best results, limit these events to no more than five. Under theseconditions, the mimics and inhibitors are stable for at least 6 months.

Table 1.
Amount per
well (nmol)

1x Resuspension Buffer to be added (μL)
for desired final concentration
2 μM Stock

10 μM Stock

20 μM Stock

0.1

50

n/an/a0.25

125*25n/a0.5

250*

50

25

1.0n/a100

50

2.0n/a200

100*this volume will exceed the capacity of a well in a 384-well plate (max 120 μL)
n/a: volumes required to resuspend at this concentration are too low for efficientreconstitution or exceed the capacity of a 96-well plate.

Plate Types and Layout
96-well miRIDIAN microRNA libraries• NUNC Polystyrene 96 well V-bottom plates (Cat #249952)• Agilent Peelable Aluminum Seal (Cat #24210-001, white) OR Thermo Scientific™ Easy Pierce™ Heat Seal (Cat #AB-3738, silver)• Default plate layout: 80‑wells per plate, columns 1 and 12 left empty“1” refers to mimic or inhibitor for microRNA 1, “2” refers to mimic or inhibitor for microRNA 2, etc.
123456789101112AEmpty

12345678910empty

BEmpty11121314151617181920

empty

CEmpty21222324252627282930

empty

DEmpty31323334353637383940

empty

EEmpty

4142

43

4445

46

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48

4950

empty

F

Empty

51

52

53

54

55

56

57

58

5960

empty

GEmpty61626364

65

66

67

68

6970

empty

HEmpty

71

7273

74

75

76

77

78

7980

empty

384-well miRIDIAN microRNA libraries• Thermo Scientific™ Abgene™ 384-Well Storage Plate (Polypropylene, Pyramidal bottom) (Cat #AB-0781)• Agilent Peelable Aluminum Seal (Cat #24210-001, white) OR Thermo Scientific™ Easy Pierce™ Heat Seal (Cat #AB-3738, silver)• Default plate layout: 280‑wells per plate, rows A and P, plus columns 1, 2 and 23, 24 left empty“1” refers to mimic or inhibitor for microRNA 1, “2” refers to mimic or inhibitor for microRNA 2, etc.
123

4

5

6

7

8

9101112

13

14

15

16

1718

1920

212223

24A Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty
B Empty Empty

1234567891011121314151617181920Empty Empty

C Empty Empty

2122232425262728

2930

3132

3334353637383940

Empty Empty

D Empty Empty

4142434445464748

49505152535455

56575859

60

Empty Empty

E Empty Empty

61626364

65

66

6768

69

70

71727374757677787980

Empty Empty

F Empty Empty

8182838485868788899091

9293949596979899

100

Empty Empty

G Empty Empty

101

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I

Empty Empty

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Empty Empty

O Empty Empty

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272273

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278279280

Empty Empty

P Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty Empty

Custom Plate Layout Requests
Catalog Libraries can only be fulfilled with the above plate layouts, since they are created from a single set of master plates. Alternative plate layouts are available forcustomer-defined Dharmacon™ Cherry-pick™ libraries, however differential pricing will apply to fulfill these requests and turnaround time will increase.

Cherry-pick Libraries
Requests for customer-specified lists of pre-designed microRNA reagents can be fulfilled directly on the Dharmacon website with the Cherry-pick Library
Plater. The addition of controls and customization of plate layout is under control of the user.
Cherry-pick Libraries can be generated for microRNA reagents from a list of any of the following identifiers:
i. miRBase ID e.g. hsa-miR-181b-5p
ii. Mature accession e.g. MIMAT0000257
iii. Dharmacon Catalog number e.g. C-311794
For questions, please contact our Technical Support staff.

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Information provided with all Library orders
Plate Maps are provided in Excel files via CD and include Sample Location (Plate and Well), Catalog Number, Mature Sanger ID, Precursor Accession Number,Mature Accession Number, and Mature Sequence.

Whole Genome Library information
Dharmacon miRIDIAN Mimic and Hairpin Inhibitor Libraries are available for human, mouse and rat microRNAs in the miRBase sequence database,version 21.0.
When unique precursors give rise to identical microRNA mature sequences, those redundant reagents are accounted for in the annotated plate map filethat accompanies the library. In this manner, the library does not contain multiple wells with identical reagents.
Mimics96-well Catalog ## Reagents# 96-well Plates

384-well Catalog ## 384-well Plates

CS-001040

2603

33CS-001040-E2

10

Mouse miRIDIAN miRNA Mimic Library

CS-002040

193926CS-002040-E2

8Rat miRIDIAN miRNA Mimic Library

CS-003040

766

11N/A

Human miRIDIAN miRNA Inhibitor Library

IH-001040

2609

33IH-001040-E2

10

Mouse miRIDIAN miRNA Inhibitor Library

IH-002040

193826IH-002040-E2

8Rat miRIDIAN miRNA Inhibitor Library

IH-003040

768

11N/A

Human miRIDIAN miRNA Mimic Library

Inhibitors

Troubleshooting
For technical information or troubleshooting contact Technical Support.

To find the contact information in your country for your technology of interest, please visit us at
horizondiscovery.com/contact-us
Horizon Discovery, 8100 Cambridge Research Park, Waterbeach, Cambridge, CB25 9TL, United Kingdom©2022 The Horizon logo and other trademarks are the property of Horizon Discovery Limited, unless otherwise stated. DHARMACON and SMARTVECTOR are trademarks ofDharmacon Inc. ABGENE, ALPS 3000, FISHERBRAND, EASY PIERCE, and DISPLACE are trademarks of Thermo Fisher Scientific, Inc.

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