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Lab Diary: Day 1

12:00 AM

1ST: SUPPLY UPDATE
My Advisor put in a special request for 1 set (=50) of RNeasy columns for me (his thesis student). I'll honestly probably need all 50, just because I have soooo many cells and I want to account for repeats.
I'm hoping it gets approved (and apparently, I'm told that it should because I'm a thesis student who needs it in order to graduate). *Fingers crossed!*

All labs have to be careful with their supplies because of the budget, so what we have right now needs to last us. Priority of supplies goes to thesis students over Work-Study and volunteers, so even though we have many new students cycling through to learn all of a sudden, unfortunately, they'll have to be content with just observing until the new funding period (since we're all trying to conserve our supplies).

Until my RNeasy columns come in (hopefully), all I can do is lots of new Westerns, which I now enjoy. All of the samples I need have already been quantified, so that's a relief!

FOR WESTERNS:
We are out of precast WB gels, so it's back to making the gels from scratch again, which is fine. I did get really good results when I made the gels myself. It takes more time, but no biggie. We are also out of 1x TGS buffer, but the recipe for making that from scratch is also pretty easy (as long as you know where everything is).

TO MAKE NEW 1X TGS BUFFER:
1L DI water
3.03 g Tris powder
14.413g Glycine powder
5mL 20% SDS

pH needs to be 8.3

NEW PRIMERS:
Last night, I found out that my Advisor told my colleague to give me specific Primers she inherited from the Thesis student before her. This was new news to me, so just now, I asked my Advisor if these primers were ones he wanted me to use so that I could read up on them. Just now, he confirmed that I will use PSA as we've been doing, and the "new" primers will be supporting primers, since they also are expressed by the Androgen Receptor. If I have "left over" cDNA, then I can use the supporting primers.

This is a great relief to me, and always very exciting, because it means I have more to work with!

CURRENTLY (when this post was first written):
Running a Western Blot from Collection #3, Samples from Day 0 (Control LNCaP) to Day 12.

I take loading accurate amounts of samples very seriously
Order of loading into wells, from L to R:
Strip-Tactin Ladder (10 uL)
18 uL of each sample:
D0
D2
D4
D6
D8
D10
D12
then 10 uL of 2X buffer to fill the remaining 2 wells so that the lanes run evenly

REALIZING:
As I'm writing this post to keep track of things in the lab, I'm reminded to quantify my "extra" days ASAP so that I can have the lysate already prepared for more Westerns, while I wait for the RNeasy columns to come in.

I meticulously collected multiple collections of 20 days all the way to 74 days, so I should be proud of showing them!


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