Wed, Aug 18 21

Received and organized VR inserts #

Most of the VR insert sequences arrived in the mail today so I spent some time resuspending those and organizing data associated with them.

Resuspension #

Resuspended all inserts in TE buffer provided by Megan (thanks!) so all inserts have a final concentration of 50 ng/ul of DNA. Then placed all inserts into a new box labeled VR fragments and placed onto my shelf in the kitchen freezer.

VR insert data management #

Cataloging #

I created this spreadsheet which catalogues each insert that has arrived, it’s total mass, and lot number.

Quality control data #

Thermo also provided a CD (???) of quality control data for each insert that arrived. I transferred that data to Google Drive but have not spent much time reviewing it.


Ran PCR with VR-7 and VR-8. VR-7 is a fragment while VR-8 is in a vector. In theory should work either way. Details of protocol reagents and primers are at this sheet. Used pFC8tac protocol as it should be suitable with these primers.

|95°C|95°C               |    |tmf:66.0
|____|_____          72°C|72°C|tmr:69.5
|5min|30s  \ 61.3°C _____|____|45s/kb
|    |      \______/ 0:30|5min|GC 55%
|    |       30s         |    |319bp

Results #

Did not have time to run out on gel; will do tomorrow. For now just nanodrop both samples results are below.

Sampleng/ul260/280260/230Sample volume (ul)

Looks like primers are working and that we can amplify small amounts of each insert in order to produce more if need be. Gel will give more complete picture.