Centrifugation

Centrifugation: RCF ↔ RPM conversion

Relative centrifugal force depends on rotor radius as well as speed, so the same rpm on two centrifuges is not the same spin. Convert either way, or rescale a protocol.

RCF ↔ RPM × g

Enter the rotor radius and either the speed or the force — the other is filled in.

RCF = 1.118 × 10⁻⁵ × r (cm) × N² (rpm)  ·  N = √( RCF / (1.118 × 10⁻⁵ × r) )

Move a protocol to a different rotor

A published method says “spin at 12 000 rpm” but does not say in what. Convert through × g so the sample sees the same force.

N₂ = N₁ × √(r₁ / r₂)

Clearing factor (k-factor) and pelleting time

The k-factor describes how quickly a rotor pellets a particle of a given sedimentation coefficient. Lower k means faster.

t (h) = k / s (svedberg)  ·  k = 2.53 × 10¹¹ × ln(rmax/rmin) / (rpm/1000)²
ParticleSedimentation coefficient
Serum albumin4.5 S
IgG7 S
Catalase11.3 S
Ribosomal subunit (bacterial, small)30 S
Ribosomal subunit (bacterial, large)50 S
Bacterial ribosome70 S
Eukaryotic ribosome80 S
Polysome (5 ribosomes)≈ 200 S
Tobacco mosaic virus185 S
Mitochondrion≈ 10 000 S

Typical spins

PurposeForceTimeNotes
Pellet bacterial cells4 000 × g10 minGentle enough to resuspend
Pellet mammalian cells200–300 × g5 minHigher force damages membranes
Clear a lysate (small scale)16 000 × g15–30 minStandard benchtop microfuge maximum
Nuclei600 × g10 min4 °C, in hypotonic buffer
Mitochondria10 000 × g15 minAfter a 600 × g pre-clear
Microsomes / membranes100 000 × g60 minUltracentrifuge required
Ribosomes150 000 × g2–3 hThrough a sucrose cushion
Exosomes100 000–120 000 × g70–90 minAfter serial pre-clears
Plasmid miniprep clearing16 000 × g10 minRoom temperature is fine

Where the constant comes from

Relative centrifugal force is the centripetal acceleration divided by standard gravity:

RCF = ω²r / g, where ω = 2πN/60 rad s⁻¹.

Substituting and expressing r in centimetres gives RCF = (2π/60)² × r/100 ÷ 9.80665 × N² = 1.11846 × 10⁻⁵ × r × N². The calculator above works in SI internally rather than using the rounded constant, so results agree with rotor manuals to four figures.

Choosing which radius to use

  • rmax — the bottom of the tube. Use this when you want to know whether something will pellet.
  • rav — halfway down the sample column. This is what most published protocols and rotor specification sheets quote.
  • rmin — the meniscus. Relevant for gradient work, where you care about the force at the top of the sample.

The difference is not trivial: a fixed-angle microfuge rotor might have rmin = 38 mm and rmax = 85 mm, so the force at the bottom of the tube is more than twice that at the top.

Common rotor radii

Rotor typermax (approx.)Max speedMax RCF
Benchtop microfuge, 24 × 1.5 mL85 mm14 000 rpm≈ 18 600 × g
Benchtop microfuge, 18 × 2 mL (compact)73 mm13 300 rpm≈ 14 400 × g
Swing-out, 4 × 100 mL180 mm4 500 rpm≈ 4 100 × g
Fixed-angle, 6 × 50 mL108 mm15 000 rpm≈ 27 200 × g
Ultracentrifuge, fixed-angle 8 × 39 mL92 mm45 000 rpm≈ 208 000 × g
Ultracentrifuge, swinging bucket 6 × 38 mL153 mm32 000 rpm≈ 175 000 × g

These are representative values only — always take the radius from your own rotor's manual.

Frequently asked questions

What is the formula for RCF?

RCF = 1.118 × 10⁻⁵ × r × N², with r the rotor radius in centimetres and N the speed in rpm. The constant comes from (2π/60)² / g expressed in cm.

Why should protocols specify × g rather than rpm?

The force a sample experiences depends on the rotor radius as well as the speed, so the same rpm on two different rotors gives different forces. Reporting × g makes a protocol reproducible on any centrifuge.

Which radius should I use — rₘᵢₙ, rₐᵥ or rₘₐₓ?

Use rₘₐₓ for pelleting (the force at the tube bottom) and rₐᵥ when comparing to published protocols, which usually quote average radius. Rotor manuals list all three.

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