how to make cluster manifolds

How To Make Cluster Manifolds

Dirt cheep vacuum manifolds with independent and disposable collection.

TSRI 3/07/2009 Brian M. Frezza

Materials

50mL Centrifuge tubes

Preferably with

smooth flat lids

The one’s pictured here come from Genesee Scientific:

Catalog #21-108TSRI 3/07/2009 Brian M. Frezza

MaterialsLarge Test Tube Rack

The size of the rack will define the maximize size of the manifold, but that

can be as large as you’d like.

Tubes should fit snugly in

the rack slots.


Materials

Make-Male Lure adaptors:Value Plastics Product #MTLCS-9

Female-Female Lure adaptors:Value Plastics Product # FTLC-6

3-way or 2-way Lure stopcocks:Value Plastics Product # VPB1000055N

You’ll need some slip style lure adaptors to connect the

junctions, and stopcocks to control the flow of vacuum. These are sold dirt cheep in

packs of 100.


Materials

You’ll need some tubing to connect everything. The diameter of the tubing must be such that it fits

snugly (air tight) around the male lures, but is strong enough not to

collapse under vacuum.

I prefer Tygon R-3603 tubing with a 1/8’’ ID

and 1/4’’ OD.

This tubing is flexible but fits tightly onto

male lures, and they’re clear so you

can see any obstructions.

You’ll also need a scissors on hand to

cut the tubing


Materials

1/4’’ Y-adaptor

Y-adaptors:Value Plastics Product #Y260-J1A

Standard Vacuum tubing that connects to a house line.

Typically 1/4’’ ID will do the trick.


Equipment

In my experience, Torx drivers work best, but a Hex (Allen) will do

in a pinch.

You’ll need a screw driver for poking lure sized holes.

The tip size must be just smaller

a male lure fitting.


Starting Checklist

• 50mL Centrifuge tubes• Male-Male lure adaptors• Female-Female lure adaptors• 2 or 3-way lure stopcocks• Flexible tubing 1/8’’ ID• Vacuum Tubing 1/4’’ ID• Y-adaptor• Scissors• Test tube rack• Screw driver


Before we get started, lets demonstrate how to use your screw driver to poke effective female

lure fittings into the centrifuge tube lids

First, remove the lid from the tube and lay it face up on a hard flat surface. Then hold the skew driver at a 90 degree angle directly above where you want to poke the hole

Next, slowly apply downward pressure to the screwdriver while holding the lid still with your other hand.

The more slowly you apply the pressure the cleaner the hole you will poke. If you go too fast, you may tear the lid around the hole. If you do, toss the lid and try again.

You’ll use this trick several times in the course of building your manifold.

Step 1: Pick a Rack Size I’m going to demonstrate using this

4x4 rack I found in a trash can.

Because that’s how I roll.

This rack in particular is really tightly packed, so we’re only going to fill

every other slot.

Also, I chose not to go with the racks the tubes are usually supplied in.

This blue rack here holds the tubes much more snugly, since you don’t want them rattling around when you

turn the vacuum on.


Step 2: Prepare the Tubes Using your expert hole poking skills learned earlier,

go ahead and poke 2 holes in each lid for the outside rows of tubes and 4 holes in each lid of the inside rows of tubes as I’ve show below.

If your rack isn’t square, arrange it so that the longer side goes left to right, and follow the same pattern as above for your holes.

Be sure to poke the holes as far apart as possible on the lids.

Long edge

2 Holes

4 Holes

4 Holes

2 Holes


Step 3: Prepare the Tubing For each tube with 2 holes, cut one

long length of tubing (~10cm)For all but one of the tubes with 4

holes, cut two short piece of tubing (~7cm)

Finally, cut 2 really long piece of tubing (~50cm) for the ends (not shown).


Step 4: Prepare the Tubing

Slide a male-male lure adaptor into each end of the all of the tubing you just cut. This should be a really tight (air tight) fit.


Step 5: Prepare the Taps Slide each stopcock into a female-female lure adaptor and then a male-male lure

adaptor as show below.

Be sure the male lure is on the bottom

Make one of these for every tube in the rack


Step 6: Prepare the Y-Adaptor

Connect the top ends of the Y

adaptor to the two really long piece of tubing as show to

the right.

Connect the bottom end of a Y adaptor to vacuum tubing large enough to

connect to a house tap as show to the

left.


Step 7: Connect the Tubes Use the small tubing to connect the 4-hole tubes in a criss-cross

pattern as show below

Use the longer tubing to connect the 2-hole tubes in column as

show below

When you’re done connecting both your rack should

look like this.


Step 8: Connect the Taps

Place one tap in each tube in the outermost hole

When you’re done that, your rack should look like this


Quick Double Check

2-way tubes should look like this (above)

4-way tubes should look like this (below).


Step 9: Connect the Y-adaptor

Connect the two ends of the Y adaptor to the end holes in the

4-hole tubes

When you’re all done, your rack should look like this.


Step 10: Connect the House Vacuum

Leave the house vacuum off when you’re not using the manifold.

Connect the other end of the Y-adaptor to the house vacuum.


Congratulations, you now have a working cluster manifold!


Independent Disposable Collection

Elutant from each tap is not mixed into a shared waste, and

instead is collected in each tube independently. This allows you to save elutant for instance when eluting your DNA from a

mini-prep.

Each collection tube is also disposable. So when you’re done collecting your sample

you can simply swap in a fresh tube and not worry about

cross-contamination.


Size Expandable

Cluster Manifold can be built to any size specification. All you need is a larger rack and more tubes.


Compare to Commercial ManifoldSigma VM20 manifold 20 tap Cluster Manifold

Standard Cluster

Collection Shared waste bin Independent Tubes

Cleaning Washing Disposable

Size Fixed Variable

Setup time ~1 min ~20 min

Cost $110.00 ~$10.00TSRI 3/07/2009 Brian M. Frezza

how to make cluster manifolds

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