Search found 5 matches

by nikinje
Wed Aug 19, 2009 4:41 pm
Forum: Cell Biology
Topic: Improving transient CHO protein expression
Replies: 0
Views: 1776

Improving transient CHO protein expression

I have started using Invitrogen's freestyle CHO cells (without serum, and in suspension) but the protein expression levels are not great compared to the HEKs I was using before. I have played around with ratios and have an optimal ratio (1:1 with 1ug DNA and 1ug MAX transfection reagent). I have als...
by nikinje
Mon May 18, 2009 2:00 pm
Forum: Molecular Biology
Topic: 96 well miniprep?
Replies: 0
Views: 1429

96 well miniprep?

I am currently trailling some 96 well miniprep kits, mainly Millipore's one and Qiagen's Turbo kit. We have done our first trial of the millipore kit and have had abysmal yields, it is a low copy number plasmid, but I was just wondering if there is anyone else out there with some experience of any 9...
by nikinje
Fri Feb 13, 2009 5:55 pm
Forum: Microbiology
Topic: E. coli Strains
Replies: 1
Views: 1877

E. coli Strains

This is a follow up to my dead E. coli problem (posted already on the forum). I have been using a kit from Clontech called in-fusion 2.0 for cloning, produced my clones and transformed them into their "super competent" FusionBlue E. coli cells. This worked fine and produced many colonies, ...
by nikinje
Wed Feb 11, 2009 2:55 pm
Forum: Microbiology
Topic: Dead E. coli problem
Replies: 2
Views: 2925

Re: Dead E. coli problem

I didn't know that about the 36 degrees, useful to know! They were put in the fridge after an overnight growup (until I could just see the colonies). I think what has happened is that they have overgrown, used up the ampicillin on the plate and thrown the plasmid out so didn't re-grow on the new pla...
by nikinje
Thu Feb 05, 2009 7:22 pm
Forum: Microbiology
Topic: Dead E. coli problem
Replies: 2
Views: 2925

Dead E. coli problem

I've had a problem recently with the E. coli cells I've been using. I transformed 7 constructs into FusionBlue competent cells (from Clontech), they grew many colonies (significantly more than my control). I performed a colony PCR 5 days later and re-streaked the colonies, but not only did the colon...