Team:MIT/Manifold
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+ | <h3 align="center" style="font-size:42px; color:teal"><b>DIY VACUUM MANIFOLD</b></h3> | ||
+ | <p align="center"><i> An Erik Ersland creation</i><br>This has been updated. It turns out brass is corroded by miniprep reagents. Do not use brass, only use plastic (preferable the same plastic for everything).</p><br><br><br> | ||
Midipreps require a vacuum manifold and minipreps may use it if you so choose. It uses vacuum to draw fluid through a spin column. Our labs manifold was in poor shape, the hose was attached with a quarter inch mass of parafilm and the opposite end from the hose leaked. A new 24 tube vacuum manifold from QIAGEN costs $270, the following kit cost 15.21(even with a lot of leftovers) and holds 36 tubes.<br /> | Midipreps require a vacuum manifold and minipreps may use it if you so choose. It uses vacuum to draw fluid through a spin column. Our labs manifold was in poor shape, the hose was attached with a quarter inch mass of parafilm and the opposite end from the hose leaked. A new 24 tube vacuum manifold from QIAGEN costs $270, the following kit cost 15.21(even with a lot of leftovers) and holds 36 tubes.<br /> | ||
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- | <a name="tools" ></a><h2>Tools | + | <a name="tools" ></a><h2>Tools</h2> |
something to cut PVC pipe, I used a hacksaw and the wood saw on my multitool.<br /> | something to cut PVC pipe, I used a hacksaw and the wood saw on my multitool.<br /> | ||
Something to tighten the brass fitting, I used a crescent wrench<br /> | Something to tighten the brass fitting, I used a crescent wrench<br /> | ||
optional: Hot glue gun or teflon tape to seal brass fitting, probably not needed<br /> | optional: Hot glue gun or teflon tape to seal brass fitting, probably not needed<br /> | ||
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- | <a name="parts" ></a><h2>Parts | + | <a name="parts" ></a><h2>Parts</h2> |
3/4" PVC pipe, the size of manifold you want determines amount. I used a 10 foot segment and probably have 4 or so feet left over. I had to cut this in half to fit it on the subway, if you have to use a train, make sure you are the type that always carries a saw.<br /> | 3/4" PVC pipe, the size of manifold you want determines amount. I used a 10 foot segment and probably have 4 or so feet left over. I had to cut this in half to fit it on the subway, if you have to use a train, make sure you are the type that always carries a saw.<br /> | ||
3X 90 degree elbows, same size as pipe<br /> | 3X 90 degree elbows, same size as pipe<br /> | ||
3X Tees, same size as pipe<br /> | 3X Tees, same size as pipe<br /> | ||
- | + | plastic barb to male thread adapter, sized for vacuum hose (I used 3/8 tube to 1/2" thread since its all I could find), the qiagen model appears to use 1/4 inch tube<br /> | |
PVC female thread to pipe slip, should fit into a tee socket and the brass fitting.<br /> | PVC female thread to pipe slip, should fit into a tee socket and the brass fitting.<br /> | ||
PVC cement<br /> | PVC cement<br /> | ||
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I bought some extras, you probably wont need any.<br /> | I bought some extras, you probably wont need any.<br /> | ||
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+ | <a name="layout" ></a><h2>Layout</h2> | ||
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I ended up making a mistake and turning the brass hose fitting 90 degrees, I think the bellow configuration is probably better for our particular setup.<br /> | I ended up making a mistake and turning the brass hose fitting 90 degrees, I think the bellow configuration is probably better for our particular setup.<br /> | ||
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<img src="https://static.igem.org/mediawiki/2014/6/67/MIT_manifold_6.jpg"><br /> | <img src="https://static.igem.org/mediawiki/2014/6/67/MIT_manifold_6.jpg"><br /> | ||
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- | Drilling the holes is the hard part. I considered lathing a piece with the correct taper and a flat on the top, then heating the pipe to make it moldable with a heat gun and pressing it into an undersized pilot hole. This is more likely to work, especially if you use a drill press and clamp the pipe to make sure its really straight. I just drilled the holes with a 5/32's drill and tilted the drill and | + | Drilling the holes is the hard part. I considered lathing a piece with the correct taper and a flat on the top (where the end is shaped like an upper case T, where the vertical part is what forms the hole and the horizontal part flattens the pipe around the hole), then heating the pipe to make it moldable with a heat gun and pressing it into an undersized pilot hole. This is more likely to work, especially if you use a drill press and clamp the pipe to make sure its really straight. I just drilled the holes with a 5/32's drill and tilted the drill and rotated it around (at about a 30 degree angle). The tilt and rotate is better than using a larger drill bit, it makes the holes conical so the top is bigger than the tube and the bottom is smaller. This angle means that you are guaranteed to get the right diameter even with imprecise holes and imprecise tubes. It also allows the tube to get wedged in tighter and seal better. Try to keep the holes circular instead of oblong, it will make them seal better. Use a piece of scrap pipe to practice and test with a spin column. Also test with a spin column regularly as you drill the final piece. Remember that you do not have a magical drill that can redrill a hole smaller, so when in doubt, drill a hole small first then re-drill bigger. Also, the spin columns wear out when you put them into and take them out of holes, so a spin column you use a lot will fit tightly in a hole that's too small for a new spin column.<br /> |
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<img src="https://static.igem.org/mediawiki/2014/4/4a/MIT_manifold_7.jpg"><br /> | <img src="https://static.igem.org/mediawiki/2014/4/4a/MIT_manifold_7.jpg"><br /> | ||
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<a name="differences" ></a><h2>Differences from Qiagen</h2> | <a name="differences" ></a><h2>Differences from Qiagen</h2> | ||
- | This manifold has 36 holes. This drains fluid into the vacuum tube, the Qiagen manifold stores fluid in the manifold and you can drain it out a capped hole on one end (the cap that leaks on ours). This means that you must have the manifold hooked up to a waste flask that is designated for QIAGEN waste (remember that mini and midiprep waste makes chlorine gas when combined with bleach). This also uses a 3/8" inner diameter hose, I think the Qiagen manifold uses something like 1/4" ID, you can probably stretch sufficiently stretchy hose.</td></tr></table> | + | This manifold has 36 holes. This manifold drains fluid into the vacuum tube, the Qiagen manifold stores fluid in the manifold and you can drain it out a capped hole on one end (the cap that leaks on ours). This means that you must have the manifold hooked up to a waste flask that is designated for QIAGEN waste (remember that mini and midiprep waste makes chlorine gas when combined with bleach). This also uses a 3/8" inner diameter hose, I think the Qiagen manifold uses something like 1/4" ID, you can probably stretch sufficiently stretchy hose, I managed to force our Qiagen hose onto this and it seals well. When adding tubes or plugs, don't force the them in. The vacuum will pull things in as far as they need to be once all holes are occupied. Forcing things into the holes may stretch them out and make the fit looser.</td></tr></table> |
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