Search found 8 matches

by biznatch
Tue Jul 27, 2010 4:01 pm
Forum: Molecular Biology
Topic: RNeasy: B-Me vs. DTT
Replies: 1
Views: 1528

RNeasy: B-Me vs. DTT

I just discovered that the QIAGEN RNeasy protocol says you can substitute DTT for B-Me in Buffer RLT for the elimination of RNases, but this only applies to isolation of RNA from animal tissue, not animal cells, yeast, plants, etc. This would be great since we use animal tissue, and it would mean we...
by biznatch
Tue Jul 08, 2008 2:18 pm
Forum: Molecular Biology
Topic: Denatured alcohol
Replies: 3
Views: 6112

Thank you! It's not that hard to come by (actually we've been using it forever and just decided to switch), but the denatured alcohol is cheaper and there's no regulations on it. E.g. with 100% EtOH we have to keep track of every time we use it to show a record when certain inspectors/safety people ...
by biznatch
Mon Jul 07, 2008 10:29 pm
Forum: Molecular Biology
Topic: primer design for DNA through mRNA sequence
Replies: 2
Views: 3570

Yes you can use the mRNA sequence to design gDNA primers. If the primers happen to span an intron (one on each side of the intron) the product from gDNA will be larger then from mRNA and if the intron is very big you might not be able to span its length by PCR. I use Primer3 (m) for designing primers.
by biznatch
Mon Jul 07, 2008 9:34 pm
Forum: Molecular Biology
Topic: Denatured alcohol
Replies: 3
Views: 6112

Denatured alcohol

Can denatured alcohol (90% ethanol, 5% methanol and 5% 2-propanol) generally be used in place of 100%/anhydrous ethanol? In QIAGEN kits it specifically says not to use denatured alcohol when making up Buffer PE but what about other things in general? Like fixing cells with 3:1 methanol:ethanol, H&am...
by biznatch
Tue Nov 20, 2007 7:37 pm
Forum: Molecular Biology
Topic: Making sodium acetate
Replies: 0
Views: 2960

Making sodium acetate

Every recipe I've found for making 1 L of 3M sodium acetate pH 5.2 goes as follows: - Dissolve 408.3 g sodium acetate-3H2O in 800 mL water. - Adjust pH to 5.2 with glacial acetic acid. - Adjust to 1 L with water. - Autoclave. I only wanted to make 100 mL so I dissolved 40.83 g in 80 mL and the volum...
by biznatch
Thu Sep 06, 2007 12:55 am
Forum: Molecular Biology
Topic: PEG c.d.
Replies: 7
Views: 4927

I too was looking for the function of PEG 8000, came across this forum topic, then the following product, and thought I'd post it back here: From USB Corporation, their PEG 8000 product page (I can't post the URL since I have less than 5 posts but Google for USB Corporation then do a Product Search ...
by biznatch
Thu Nov 16, 2006 4:56 pm
Forum: Bioinformatics
Topic: Want to identify common nucleotide motifs
Replies: 5
Views: 5886

Thanks, I've tried MEME but ya it has a maximum of 60kb for all the sequences. I'm not really sure what I'm looking for, just doing a screen for a group of genes I've found to be commonly regulated and want to find possible common motifs in their genomic sequences. I'd like to compare the gene itsel...
by biznatch
Thu Nov 16, 2006 5:59 am
Forum: Bioinformatics
Topic: Want to identify common nucleotide motifs
Replies: 5
Views: 5886

Want to identify common nucleotide motifs

I'm looking for a program/website that can identify motifs common between a number of long nucleotide sequences. I'd like to compare sequences (the more the better, 5-10 would be the minimum) each up to 150kb. I've found programs for 2 long sequences, or lots of short ones, but nothing for several l...