Open House preparation

VDSers, to get ready for the Open House this week. I would like for each of your to walk through the GOLD/PyMol demonstration. Here is a link to the Instructions on Google Docs.

This should be done by Tuesday so that I can put things together.

https://docs.google.com/fileview?id=0B_Gl3lMyhDsoMzA1YTUwM2ItNzQ4OS00ZDFlLWFmMDItYTIxYTNhZDQ5OTVm&hl=en&authkey=CI23g_EM

Also, each person will need to make a slide or two of their research. A template has been uploaded for you to use.

https://docs.google.com/fileview?id=0B_Gl3lMyhDsoNDI5MzcwYzgtOWU4Yy00ZmQ2LTg3NzAtZmEwZjM5N2JiZGM4&hl=en&authkey=CPPfzPoO

Once you have finished your slides – upload them to the GoogleDocs folder – OpenHouse.

Running Gels after PCR for pNIC-Bsa4

Zoe asked:

I am coming in to make my gel and run the samples. I was looking at the protocol, and it says “Use 100kb ladder as marker” Then it says right under that 100 bp ladder. Which should we use? How much blue juice should I put in each sample (2 uL?) ?  How long do we need to run the gel for and how far down should our samples be?

Thank you,

Zoe

Dr. B says:

Yes – TYPO – should be 100 bp ladder.

amount of blue juice depends on the volume of your sample.
Remember to only take HALF of you PCR sample.
So that should be 12.5 ul if you started with 25 ul.
Then BLUE JUICE is 6X concentration stock and you want it to be around 1X final in the tube..
So, 12.5 divided by 6 is roughly 2 ul.

The blue dye front of you gel should be at least 3/4 th of the way down the gel.

Dr. B

Labeling plasmid DNA stocks

After midiprep or maxiprep, you want to label your tubes of stock DNA well because you spent a lot of time and effort making it. Also, this DNA will be shared by the whole lab so it should be well labeled.

Label the top and side of tube – use the red sticky dots if necessary (the white squares are lame)

Put:      Initials, Date, type of DNA (pNIC-Bsa4, pGBR22, HSC0041026, etc. ), the concentration in λ ( lambda = micrograms/microliter )    OR  put the concentration  in nanograms/microliter. If it contains a gene inside of a vector – you should put the Gene name and the Vector name (e.g.     DHFR in pNIC-Bsa4)

Target Assignments

Target Assignments for Returning VDSers. Need to Submit your plasmid to DNA sequencing to determine what it is.

HsCD00041026 – Thao

HsCD00002513 – Candace

Zhang – Mike

HsCD00040190 – Da’Marcus & Zoe