Search found 13 matches

by jamiejlg
Fri Dec 21, 2007 8:02 pm
Forum: Cell Biology
Topic: yeast Media
Replies: 2
Views: 2315

Hi- thanks for your reply...I never did find out what was the cause of the precipitation. I just opted to sterile filter the media instead and save myself the grief! Thanks and happy holiday!!
by jamiejlg
Mon Dec 17, 2007 7:45 pm
Forum: Cell Biology
Topic: yeast Media
Replies: 2
Views: 2315

yeast Media

Hi folks! I have a mysterious media making occurrance- hope you can help... I am making Semi Rich Media (SRM) for yeast. SRM is : 0.2% Bacto peptone 0.1% yeast extract 0.3% casamino acids 0.17% YNB without amino acids and ammonium sulfate 0.06% KH2PO4 1% glycerol I adjust the pH to 6 with 2M NaOH an...
by jamiejlg
Wed Jul 11, 2007 6:45 pm
Forum: Bioinformatics
Topic: Which sequence assembly program??
Replies: 2
Views: 4095

Which sequence assembly program??

There are so many...I just need to do simple contig alignments like AutoAssembler used to do, and I'm finding Bioedit to be cumbersome. Does anyone know of a program similar to AutoAssembler? Do I really have to buy Sequencher?
by jamiejlg
Wed Jul 11, 2007 5:22 pm
Forum: Molecular Biology
Topic: cloning troubles
Replies: 5
Views: 2936

just an update for anyone who might have wondered and the nice people who offered suggestions. I was able to subclone my 230 bp gene fragment with Invitrogen's zero-blunt kit and then insert it into my desired vector. I did a side by side sequence verification with T7 endonuclease treated and withou...
by jamiejlg
Wed Jul 11, 2007 4:22 pm
Forum: Bioinformatics
Topic: Bioedit alignment
Replies: 1
Views: 8627

Bioedit alignment

Hi- I'm using BioEdit for the first time to do a simple alignment but having trouble. It's just a forward, reverse and respective primers.
Do I need to complement my sequences or does bioedit (clustal, really) automatically do it?

Thanks,
Jamie
by jamiejlg
Wed Jul 11, 2007 4:15 pm
Forum: Molecular Biology
Topic: Bioedit Sequence Assembly
Replies: 1
Views: 1778

Bioedit Sequence Assembly

Hi everyone,
I am trying to analyze some sequence data using Bioedit, but the sequences do not fit together. It's only a simple alignment- a forward and reverse and the respective primers. It should go together easy.

Does anyone know if Bioedit (Clustal, really) automatically reverse complements?
by jamiejlg
Sun May 13, 2007 1:59 pm
Forum: Molecular Biology
Topic: MJ PCR machine PT200 gradient question
Replies: 4
Views: 3296

good question- I didn't ask, but should be speaking with him shortly and I'll let you know what I find out-
by jamiejlg
Sun May 13, 2007 1:53 pm
Forum: Molecular Biology
Topic: cloning troubles
Replies: 5
Views: 2936

cloning troubles

Thanks for your answers from my last post. I'm having trouble with a cloning that shouldn't be so difficult, but I'm not getting transformants. Do any of you have suggestions for the following? I'm synthesizing a 230 bp fragment through PCR using overlapping oligos. I got a PCR product that appears ...
by jamiejlg
Thu May 10, 2007 7:20 pm
Forum: Molecular Biology
Topic: analyzing a ligation before transformation
Replies: 8
Views: 15414

analyzing a ligation before transformation

is it standard practice to run a ligation reaction on an agarose gel to determine if the pieces were ligated before proceeding to transformation?
by jamiejlg
Sat Apr 14, 2007 2:55 pm
Forum: Molecular Biology
Topic: MJ PCR machine PT200 gradient question
Replies: 4
Views: 3296

Hi- thanks for your reply. I contacted a rep- the machine I have is a gradient, but the manual just shows the gradient along the 12 well side. According to the rep- for the split block, the gradient runs along the 8 well side. Hopefully, anyway- since that's what I'm going to try!
by jamiejlg
Fri Apr 13, 2007 8:23 pm
Forum: Molecular Biology
Topic: MJ PCR machine PT200 gradient question
Replies: 4
Views: 3296

MJ PCR machine PT200 gradient question

Hi there,
I'm working in a new lab and they have the MJ research PT200 thermocycler. The dual block holds 48 0.2 tubes on each side, 6 across and 8 down. Does anyone know the orientation of the block so I can set up a gradient? The manual doesn't show the dual 48 well block.
Thanks,
Jamie
by jamiejlg
Tue Feb 27, 2007 6:37 pm
Forum: Molecular Biology
Topic: cloning 5kb insert
Replies: 2
Views: 1649

cloning 5kb insert

Hello, I have to clone a gene that is nearly 5kb. Can anyone suggest a specific vector that can handle larger fragments?