Search found 90 matches

by oppox
Thu Aug 17, 2006 11:33 pm
Forum: Molecular Biology
Topic: Primer design
Replies: 2
Views: 1747

I have never calculated any delta G values, just calculating the Tm. I have designed primers 50 bases long and I think they can be longer, they should atleast be around 20-25. The disadvantages I can think of is maybe its harder to synthesize long primers and u may have some unspecific binding perha...
by oppox
Mon Jul 24, 2006 1:11 pm
Forum: Physiology
Topic: Sex and emotions
Replies: 4
Views: 6428

uh well from my own experiance I know that what u say isnt true.

I think its a social thing more than hormones, more often guys dont show their feeling and their guy gang probably aint the best emotional forum they can find.
by oppox
Mon Jul 24, 2006 1:01 pm
Forum: Molecular Biology
Topic: Molecular Biology Lab Techniques - Significance
Replies: 3
Views: 1946

thee are alot of standard ways of making and analyzing things, start by looking them up. Its quite impossible to write tips on how to analyze proteins since there are so many different proteins and different methods on how to analyze them. If you are stuck on something specific im glad to see if my ...
by oppox
Tue Jul 11, 2006 9:55 pm
Forum: Molecular Biology
Topic: kanamycin resistance problems.
Replies: 7
Views: 4470

hmm its something funky about this, I made fresh never frozen Kanamycin today and I did a new conc test with untransformed bl21 cells. Hope I can see some results tomorrow, thx for the help.
by oppox
Tue Jul 11, 2006 8:20 pm
Forum: Molecular Biology
Topic: How do primers work?
Replies: 5
Views: 27793

The primers are as u say a short designed complementary sequence, they come in pair, one forward (complementary to the non coding strand) and the reverse primer (complementary to the coding strand). After the first heating step, the DNA separates and when the temp lowers down the primers anneal, for...
by oppox
Tue Jul 11, 2006 4:26 pm
Forum: Molecular Biology
Topic: kanamycin resistance problems.
Replies: 7
Views: 4470

So not freezing the kan solution? ok.
In novagens manual, the working concentration of Kan is 30.
Your cells do not grow untransformed?
thx for the help.
by oppox
Tue Jul 11, 2006 1:13 pm
Forum: Molecular Biology
Topic: kanamycin resistance problems.
Replies: 7
Views: 4470

kanamycin resistance problems.

Im trying to get a pET28 vector into a cell (any cell, ive tried most of them) the vector has Kan resistance. The problem though is that the cells without vector grow pretty good on Agar-Kan plates, so I cant destinguish between positive cells. Ive tried to screen with different concentrations of ka...
by oppox
Sat Jul 01, 2006 10:52 pm
Forum: Molecular Biology
Topic: DNA denaturation during PCR ??
Replies: 5
Views: 2634

well its a way, but an ordinary pcr machine cant flash cool the sample. If u mix DNA and primers heat it to 95 and the transfer it to dry ice or something u get the effect, but it takes quite some time if use a non robotic pcr machine to cool it to the annealing temp. Im not an expert on this but I ...
by oppox
Sat Jul 01, 2006 10:38 pm
Forum: Molecular Biology
Topic: smear for PCR products
Replies: 8
Views: 15768

yea begin to dilute it like 50 or 100 times or even more
by oppox
Fri Jun 30, 2006 10:11 pm
Forum: Molecular Biology
Topic: DNA denaturation during PCR ??
Replies: 5
Views: 2634

without primers they would but u got so much more primers then template so its just a matter of probability
by oppox
Fri Jun 30, 2006 2:14 pm
Forum: Molecular Biology
Topic: single stranded cDNA storage
Replies: 6
Views: 22666

Cant u ask the ones who told you what to do. Singlestranded should be more unstable than double, I dont think storing it at -80 will hurt.
by oppox
Fri Jun 30, 2006 7:44 am
Forum: Molecular Biology
Topic: MAINTANENCE AND EXPRESSION HOST CELLS
Replies: 1
Views: 1289

I have never come across top10 before, but a little search in google it turned up to be from invitrogen and from what I could understand they are kind of the same as xl1blue cells.
http://www.invitrogen.com search for top10.

Bl21, doesnt produce alot of plasmids but it expresses genes and produces protein.