Recent content by Goodie

The polyA is upstream of the promoter for the reporter gene Can you explain more of this? Where did you get this? Thanks.

I get different answers in this issue which makes me confused. :confused: So again, should i leave the UV light 24/7 when i don't work? Should i turn the blowers off when i have the UV light on? Should i have the blowers on when i work? Should i turn the blowers off when i don't...

Usually the poly A sequence lays downstream of a gene to give it good RNA termination and translation. BUt when it lies upstream is something new :bugeye: . Upstream means before the promoter. How come?

# About the laminar flow hood; when should i turn off the blowers which circulate the air? After the work or should it be on all the time? Can i have it on at the same time with the UV light? :frown:

What is the difference between mold, fungi, yeast and bacteria? :rolleyes:

• Why should some Poly-A sequences lay upstream of a reporter gene? • Isogenic cell lines? • Null cell lines? Any ideas are greatly appreciated.

# Do I need mRNA destabilization signals for my gene expression? If so where should I add it? # It says; "cDNA are often obtained by addition of homopolymeric tails into the 5’ non coding region which should be removed.” Which homopolymeric tails are usually used for this purpose? # Which...

As i know RT-PCR means Real Time PCR and Reverse Transcription PCR. When i want to examine up or down regulation of my gene expression which one should i use; Real Time PCR or Reverse Transcription PCR? :eek:

So does it mean that a 80-90% confluence is most probably at mid log phase and 100% confluent is at end log phase? What do you mean? For example why one splits a flask into two new flasks (split ratio 1/2) instead of 4 new flasks (1/4) ? Is that the amount of cells one needs that also decide...

So when my flask is confluent then my cells are at the mid log phase? The split ratio is usually 1/2 to 1/20. What decides which factor we should use? :rolleyes:

Should I always split my cells at their mid log phase? Or does it only matter at transfection? How do I know that my cells are in the mid log phase? How do i know that my cells are 80-100% confluence? Is this 80-100% a rough estimation or accurately done? Thanks.

links and info of RT- PCR? appreciate for any inputs. :cool:

anyone with good links of Microarrays? thank you.

if a cell line has a maximum of 10 passages before it changes character and i only have done 4 passages of it and made the frozen stock of some of the cells. how many passages does this frozen stock have if i thaw it and passage it? will it be only 6 passages left or 10 passages? thanks for...

Thanks alot. :smile: