Questions relating to protocols, procedures and good practice when using laboratory equipment
6
votes
1answer
62 views
What is the best way to express two proteins in a mammalian cell?
I have two proteins and I will be preparing a vector with both genes for stable transfection. Each protein will have their own promoter and I will use piggyBac vector to insert a single cassette with ...
5
votes
1answer
107 views
Using ion-exchange chromatography to purify DNA from a cell extract - Is DNA more negatively charged then RNA?
When applying this method we have a glass or plastic column of resin which is positively charged. Then we pour cell extract into the column in order to capture the negatively charged particles which ...
1
vote
1answer
25 views
How can I save bacillus strains on filter paper without an -80 degree freezer?
I want to save my bacillus strains but I don't have access to a -80 degree freezer. What are possible alternatives?
3
votes
0answers
22 views
Can I purify polyhydroxyalkanoates by heating the cells extensively?
Traditional methods of purifying polyhydroxyalkanoates (PHAs) and other bioplastics made by bacteria involve washing the cells with harsh chemicals or strong bases.I'm interested in maintaining the ...
2
votes
0answers
28 views
How long does Lentivirus take to express in vivo mouse neurons?
Does anyone know how long it takes for a standard Lentivirus vector to express its genes (under a strong promoter such as CAG, CB7, etc.), after injection into the brain of a mouse?
By hearsay I ...
2
votes
0answers
22 views
How do you store membrane proteins?
We're producing some membrane proteins and they aren't amenable to freeze thaws even when we add glycerol. The proteins are solubilized in detergent above the cmc so they should be in micelle form in ...
2
votes
0answers
30 views
Does preservation in ethanol alter leaf litter mass?
I have benthic samples that were collected with an Ekman dredge from some small ponds. The samples contain quite a bit of coarse particulate organic matter (CPOM, basically dead leaves). I would ...
2
votes
0answers
39 views
Resuspending Cells from a filter plate
My experiment involves pulling down cells on to a filter plate for a variety of assays. However, I want to ensure that my cells are alive so I am trying to resuspend them so that I can do an accurate ...
2
votes
0answers
15 views
Effect of Ethanol on Brain Volume Measurements
I want to compare the brain sizes of two populations of fish. However, all the samples I have were fixed in 95% ethanol. As far as I know, 4% formalin is the normal fixative for soft tissues. Can I ...
1
vote
0answers
29 views
How can I view modENCODE data faster?
I am trying to view several data tracks in the modENCODE GBrowse genomic browser. However, the site is so slow, it is practically unworkable. Is there a faster way to explore the data?
1
vote
0answers
30 views
How is a blood sample analyzed to give the different levels of metabolites?
How is a blood sample analyzed to give the different levels of metabolites?
Are there any any procedural differences with each blood test?
Thanks in advance!
1
vote
0answers
32 views
Can I heat Trizol?
I wonder if I can heat Trizol reagent for 30 min 65C. The goal is to disrupt protein-RNA complex while inhibiting nucleases. (I can't use RNasin cause it's inactivated in 65C, and can't use RVC cause ...
0
votes
0answers
20 views
What material is recommended for autoradiography cassettes when working with 32P?
So, during radiation safety training, much hullabaloo is made regarding how the high energy beta emitted by 32P produces Bremsstrahlung radiation when striking a high Z shielding material. As such, ...
0
votes
0answers
17 views
Western blotting of lung samples
I have been working on lung samples to perform western blotting. I have two related questions about this:
I get very nonspecific results: at times nothing at all, at times it glows on membrane but ...
