19 replies to this topic

[medchemgirl]

SB works better when resolving really small DNA fragments. You can even see differences in migrations from 2 or 3 bp in difference.

swanny, on Jan 27 2009, 10:39 PM, said:

clone, on Jan 6 2005, 06:09 AM, said:

Hi bob1
thabx alot for the SB buffer information. I used it but i m facing problems in resolution. I run 1kb marker on 0.8% agarose with 250volts for 30 min .
please help me if u have better results
regards

I have been able to resolve the two ~500 bp bands in a size ladder. Instead of NaOH and boric acid, I use sodium tetraborate and boric acid, which is in the Biotechniques paper.

The gels run fast, but there is no real buffering capacity, so you have to replace the buffer fairly regularly. I have tied recovering it and reusing it, but that's only good for a few runs. haven't tried 'spiking' the old buffer with fresh buffer occasionally, though, but it might work if you're really pressed for money.

[medchemgirl]

I cut prices by getting as many samples and reagents for free as possible. Last year I got about $3,000 in equipment and reagents for electrophoresis and 2D electrophoresis from Invitrogen. I got gel boxes for mini gels and midi gels, and the transfer box for western blot. I got precast gels, the buffers, the loading buffers, everything for free. These were new products they wanted to try and sometimes they are willing to give it for free so that they can get a feedback in the equipment. Just seek out, talk to people, fill out surveys and look for free stuff. Really, about 1/3 of the stuff I use I've been able to get for free. It makes your boss really happy.

[massiveattack]

Hi,

In our lab we also pack our own tip boxes for non sterile purposes whilst buying barrier tips for molecular biology procedures. We even use glass pipettes for cell culture which we wash and bake rather than buying the plastic ones. We make and filter our own media (it's cheaper to buy the media powder form in bulk and make your own media than ordering indiviual bottles from the same company). As the above poster mentioned, company freebies are a good way of getting by when on a tight budget (one of the students in my lab pretty much did her phD experiments using freebies such as sample vectors, kits etc). Our lab doesn't use much kits, we often do things the old fashioned way, for example, we make up our own solutions for cloning. The most important advice the students of our lab are often told is to plan experiments well and always be aware of what one is doing so as to minimise errors and repeating of experiments, and hence wasting of reagents!

Cheers,
massiveattack

[lab rat]

labrat, on 29 January 2005 - 09:21 PM, said:

Here is another way of cutting lab cost - to reuse TAE buffer and even reuse the gel.
http://www.protocol-online.org/forums/inde...?showtopic=4972

We reuse our polyacrylamide gels several times. I typically use mine 2-3 times a day, and make new every week. We put EtBr in the running buffer, take pix, and run the gel an hour or so before loading the next run. We use TBE, but I think I'll try bob's suggestion this week. Thanks bob!

Buying the pre-cast glass for sequencing gels can get expensive. Go to the local window glass shop and take one of the cracked originals with you. Ask to have them cut a replacement, which cost us about $15/plate. They can cut the ears, too; tell them to drill a pilot hole in each corner before making the cuts to avoid breaking the glass.

Rain-Ex (car window water repellant) is the same as Gel-Repel, and can be purchased much more cheaply. If you don't need to transfer the gels, then you can skip treating the glass.

A fish tank thermometer strip is an alternative to gel strips.

Use isopropyl alcohol instead of ethanol to precipitate DNA. You only need to add 2/3 isopropanol to the aqueous layer and put in the freezer for an hour before centrifugation. This means you can do extractions in 1.5 ml tube, so you only use 3 tubes to do a single CTAB-chloroform extraction. (CTAB/boil, chloroform, aqueous solution/precipitation and storage)

[Trof]

I recently cut the cost of our sequencing mix by 3/4, but that's probably no big deal, because everyone under financial pressure already did that.
We used standard protocol from ABI (BigDye master mix), 4 ul of mix, 2 ul of sequencing buffer in 20 ul reaction.

I learned on the workshop that this could be done using only half in half a volume.
So I tested several variants, and ended with using 1 ul of mix, 1.5 ul of buffer in 10 ul volume (you need good tightly capped tubes though), which uses only 1/4 of original mix for the same reaction. There is no difference in the results and no changes in run program. It even cuts the cost of purification mix (XTerminator) by half due to the reduced volume, but purification can be made also by cheap ethanol method without any kit, just that the kit gives cleaner beginning reads.

I found a page about plasmid sequencing, that went really extreme in diluting the mix, but I don't thing I'd like to go that far by increasing the number of cycles or so.