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How much to order

The size to order for one knockdown experiment

GuidelinesProtocols
01 Select product type
02 Medium in the well
03 Wells per sequence

A titration at 5 μM, 10 μM and 20 μM in triplicate is 9 wells.

9 wells: 3 at each of 5 μM, 10 μM and 20 μM.

04 The cells

1 dose: 3 days is within one re-dosing interval.

Show every combination

The tables show the whole wells one vial gives at each concentration, one treatment each. Sizes above 100 nmol are sold by quotation.

Each well of a 96-well plate holds 100 μL
Size5 μM10 μM20 μM
10 nmol20105
25 nmol502512
50 nmol1005025
100 nmol20010050
Each well of a 48-well plate holds 250 μL
Size5 μM10 μM20 μM
10 nmol842
25 nmol20105
50 nmol402010
100 nmol804020
Each well of your 24-well plate holds 500 μL
Size5 μM10 μM20 μM
10 nmol421
25 nmol1052
50 nmol20105
100 nmol (the size to order)402010
Each well of a 12-well plate holds 1 mL
Size5 μM10 μM20 μM
10 nmol210
25 nmol521
50 nmol1052
100 nmol20105
Each well of a 6-well plate holds 2 mL
Size5 μM10 μM20 μM
10 nmol100
25 nmol210
50 nmol521
100 nmol1052
Adjust the wells

Multiplies the wells per sequence.

Multiplies the wells per sequence.

The 24-well plate holds 500 μL.

Show the dilution

The working solution at 10 μM from the 1 mM stock, in complete medium.

For a single well:

  1. Step 01

    Add stock solution

    5 μL

    of the 1 mM stock

  2. Step 02

    Add buffer

    495 μL

    complete medium

  3. Step 03

    Final solution

    10 μM

    500 μL (1:100 dilution)

For 10 wells (with 20% excess):

Mix 60 μL stock + 5,940 μL buffer = 6 mL of working solution

In the same well, 2.5 μL of stock makes 5 μM and 10 μL of stock makes 20 μM.

Compare the products
Self-delivering compared with transfection-optimized
FeaturesdASOtoASO
Transfection requiredNoYes
Typical concentration5-20 μM50-100 nM

Guidelines sdASO and toASO

ASO usage guide

sdASO

AUMsilence sdASO guidelines

Getting started

The recommended working range is 5-20 μM, with a starting concentration of 10 μM. The optimal concentration varies with the target gene, the RNA class (messenger RNA, microRNA or long non-coding RNA) and the cell type, and should be determined by titration for each system.

The same range for every cell type

From primary neurons and cardiomyocytes to immortalized lines and fast-dividing cancer lines, the range above is the one to work in. Where inside it a particular line lands is what the first experiment tells you.

Protocol overview

  1. 01Reconstitute lyophilized ASO to 1 mM in nuclease-free water
  2. 02Aliquot and store at -20°C or -80°C
  3. 03Dilute to working concentration (5-20 μM) in complete media
  4. 04Add directly to cells: no transfection reagent needed
  5. 05Incubate for 24-72 hours
  6. 06Assess knockdown efficiency by qRT-PCR or Western blot

Key advantages

  • No transfection reagent required
  • Reduced toxicity compared to transfection
  • Works in serum-containing media
  • Suitable for difficult-to-transfect cells

Optimization strategy

  1. 01Start at 10 μM
  2. 02Assess knockdown at 24-72 hours post-treatment
  3. 03If knockdown is poor: move up towards 20 μM
    If knockdown is strong: move down towards 5 μM

toASO

AUMsilence toASO guidelines

Working concentration

Final working concentration: 50-100 nM

This range holds for every transfection method.

Transfection methods

Transfection methods, their key features and what each is best for
MethodKey featuresBest for
Lipid-basedMost common, gentleMost adherent cell lines
ElectroporationHigh efficiency, no reagentPrimary cells, suspension cells
Calcium phosphateCost-effectiveSome adherent cells
Polymer-basedLow toxicitySensitive cells

Lipid transfection protocol

  1. 01Prepare cells at 50-70% confluency
  2. 02Dilute toASO in serum-free medium
  3. 03Dilute transfection reagent separately
  4. 04Combine and incubate 5-20 min
  5. 05Add complex to cells
  6. 06Assess knockdown at 24-72h post-transfection

Troubleshooting guide

Low knockdown?

  • Move up within 50 nM to 100 nM
  • Optimize ASO:reagent ratio
  • Try a different transfection method

High toxicity?

  • Reduce concentration toward 50 nM, the bottom of the range
  • Decrease transfection reagent
  • Shorten incubation time

Coverage calculator

Number of wells per product size (96-well plate at 50 nM, 120 μL in the well with the two dilutions):

  • 2 nmol = 333 wells
  • 5 nmol = 833 wells
  • 10 nmol = 1,666 wells
  • 25 nmol = 4,166 wells

Actual coverage depends on confluency, controls, and safety margins.

Both products

General best practices

Storage & stock preparation

  • Store lyophilized ASOs at -20°C (stable for years)
  • sdASO Standard stock: 1 mM
    • What the protocol tables are worked from: 1% of a well's volume at 10 μM, a minimal addition to the cells
  • toASO Working stock: 2 μM
    • Pre-diluted for direct use in transfection
    • Minimizes pipetting errors
  • Aliquot to avoid freeze-thaw cycles (≤3 cycles)
  • Store aliquots: -20°C (1 month) or -80°C (6 months)
  • Working solutions stable 48h at 4°C

Experimental controls

  • Negative control: Scrambled sequence ASO
  • Positive control: Validated ASO (e.g., GAPDH)
  • Mock control: Transfection reagent only
  • Untreated control: Cells with media only

Validation methods

  • qRT-PCR: The most commonly used method for mRNA quantification
  • Western blot: Confirmation of protein knockdown
  • Flow cytometry: Cell surface proteins
  • Immunofluorescence: Protein levels

Timeline considerations

  • mRNA knockdown: 24-72 hours
  • Protein knockdown: 48-96 hours
  • Phenotypic changes: 72+ hours
  • Re-dosing: Every 3 to 5 days if needed

For research use only. Not for use in diagnostic or therapeutic procedures.