I've heard of matrigel, although I guess I would have to order a kit for that? isn't it expensive? Aren't micro-boyden chambers used to study chemokine effects on leukocytes? I just want to test the effects of chemotherapy drugs on the cell motility and invasiveness...can boyden chambers be used for...

Hello experts, I am doing some tissue culture work with cancer cells and was wondering if anyone knew of any quick and simple methods to measure cell motility or invasiveness? A lot of the assays for cell motility seem too time consuming. As a last resort, I suppose there are kits out there that ser...

Hi all, I've been having a great deal of problem transferring my proteins over to my nitrocellulose membrane for western blots. The strange thing here is that my ladder has actually completely transferred over and even passed through the membrane while my protein samples are all mostly stuck onto th...

Hi all, I am doing some QPCR experiments with SYBR Green using cDNA and my primers never seem to work properly. I re-designed it 3 times already, the first time the primers looked promising- from the melting curve there were no primer dimers but then the CT values were flutuating up and down for my ...

Hi canalon,

Yea, I was a bit afraid of my low concentration of DNA. I will re-do the transformation then! Thanks.

Hi all, I was doing some plating on LB plates and let it grow overnight. The next day I noticed that I had a lawn of bacterial growth, probably becuase I plated the entire transformation mix and it was way too concentrated. Anyhow, I was just wondering whether it would be ok to replate from this bac...

I am transforming into DH5alpha competent cells, I think the ligation mix is alright...I'm trying it with fresh buffer and ligase in a 20ul ligation mix at 22 degrees for an hour now at various molar ratios. I re-made a fresh batch of insert and vector both around 30ng/ul. If I used 1ul of both inse...

O you are right! I didn't even about that yet...it would be hard to see but as of now I'm having enough trouble just getting some colonies...one step at a time I guess...

My band does migrate to the expected size but I know if I can call it clean since there is a smear...but my uncut vector doesn't have a smear though

Hi everyone, I was wondering why I keep seeing a smear in my bands after digestion of my vector. I digested with 1ul EcoR1 and XhoI in a 50ul reaction for 2-3 hours. I also ran the uncut vector along side it and noticed that the smear appears only after digestion. My post-doc told me to just cut the...

Alright I will try it out...but if I dilute my vector I will need even less of the insert right which means less volume...but I'm trying to increase the volume of the insert while keeping the volume of the vector around 1ul...I guess I can always dilute my insert as well...we will see...

So, using 1ul insert and vector has worked for you? Is it in a 20ul ligation reaction? Do you mind if I ask what concentration of vector and insert you start out with before and after purification?