Search found 259 matches

by biology_06er
Mon Oct 13, 2014 11:37 pm
Forum: Molecular Biology
Topic: Flow cytometry
Replies: 0
Views: 1616

Flow cytometry

Hi there, So I have carried out flow three times now to measure fluorescence (each time one sample per what I was looking at i.e no triplicates or anything)...and each of the three times the exp was carried out the same way. Would it be OK to combine the fluorescence of three exp, get average/standa...
by biology_06er
Sun Jun 16, 2013 12:15 pm
Forum: Microbiology
Topic: Getting Bacteria from an OD0.8 to an OD 0.1
Replies: 1
Views: 2200

Getting Bacteria from an OD0.8 to an OD 0.1

Hi there, Say I have 50mL of bacteria at an OD0.8 and I need to get bacteria to a reading of 0.1..this is what I've been doing in the past but just want to confirm that what I've been doing in correct. So from my 50mL, I took 1mL, spun and then resuspended the pellet in 1mL of PBS. (I now have 1mL o...
by biology_06er
Fri Mar 29, 2013 1:04 am
Forum: General Discussion
Topic: Plasimd prep from frozen cells
Replies: 1
Views: 1629

Plasimd prep from frozen cells

Hi there,

I read in a protocol that that it is possible to extract plasmid from frozen cells using a kit? How would one do use frozen cells? Would you take a glycerol stock, thaw it out, spin it and use pellet?
by biology_06er
Wed Feb 27, 2013 3:38 am
Forum: General Discussion
Topic: TCA preipitation
Replies: 2
Views: 4687

OHHH, do i TCA the s/n from step 3???
by biology_06er
Wed Feb 27, 2013 3:25 am
Forum: General Discussion
Topic: TCA preipitation
Replies: 2
Views: 4687

TCA preipitation

Hi there, I have to do a cell wall extraction on bacteria soon and my protein of interest is cell wall anchored. My supervisor told me to do TCA precipitation (haven't done before) but I assume it's too isolate the proteins within the cell wall?? He told me I would have three components by the end o...
by biology_06er
Sun Jul 01, 2012 11:55 am
Forum: General Discussion
Topic: Resuspending bacterial pellet to a certain OD600
Replies: 8
Views: 5905

and from that 1mL resuspended culture..I take

0.5*2mL/1.7=588uL and top up to 2mL
by biology_06er
Sun Jul 01, 2012 11:53 am
Forum: General Discussion
Topic: Resuspending bacterial pellet to a certain OD600
Replies: 8
Views: 5905

Re: Resuspending bacterial pellet to a certain OD600

Urg..I meant resuspened in 1mL
by biology_06er
Fri Jun 29, 2012 10:08 pm
Forum: General Discussion
Topic: Resuspending bacterial pellet to a certain OD600
Replies: 8
Views: 5905

That sounds wrong.. Is this way better..say I had the a 10mL of o/n starter culture I take 1mL from that starter culture, wash, spin, block etc..and when it comes down to resuspending my pellet..i resuspend in 1.5mL again..therefore I technically have a culture at an OD600 of 1.7. From that culture ...
by biology_06er
Fri Jun 29, 2012 9:54 pm
Forum: General Discussion
Topic: Resuspending bacterial pellet to a certain OD600
Replies: 8
Views: 5905

Re: Resuspending bacterial pellet to a certain OD600

Something like that?..Which means a part is wrong?
by biology_06er
Fri Jun 29, 2012 10:00 am
Forum: General Discussion
Topic: Resuspending bacterial pellet to a certain OD600
Replies: 8
Views: 5905

Re: Resuspending bacterial pellet to a certain OD600

As in from my o.n culture take 0.5/1.7=0.29*2mL=0.588..So do I take 588uL of my overnight culture..spin it down wash etc etc..then resuspend the pellet in 2mL?
by biology_06er
Fri Jun 29, 2012 2:23 am
Forum: General Discussion
Topic: Resuspending bacterial pellet to a certain OD600
Replies: 8
Views: 5905

Resuspending bacterial pellet to a certain OD600

Hi there, I plan to do some flow next week and an one of the steps is to resuspend the bacterial pellet to an OD600 0.5. So say I had a 1.5mL overnight culture and the following morning the OD600 was 1.7. Once I spin down, wash etc etc I read to get to a certain OD (0.5 in my case)...this is the cal...
by biology_06er
Wed Jun 27, 2012 10:19 pm
Forum: Molecular Biology
Topic: GFP fusion protein not expressing!
Replies: 1
Views: 1906

GFP fusion protein not expressing!

Hi there, I tried to make a GFP fusion protein (my gene of interest was fused to the C terminal of GFP) but no expression occured...I tested the expression of the two proteins alone and individually they worked but not when fused together..I tried a linker between the two..still no luck...could it j...