User:Anthony Salvagno/Notebook/Research/2010/06/22/Tethering Planning Notes

Tethering sucks but here is a bunch of things I need to do to get tethering working:

Things I've Tried
Brian is running the PCR to make more 4.4kb stretch DNA.
 * washed invitrogen fluospheres to remove streptavidin, resuspend in BGB
 * concetrated anti-dig
 * incubate beads with DNA
 * high salt beads during tethering, wash with BGB

To Try
For tethering: Other:
 * Digest lots of pBR322
 * extract pBR322 and purify
 * Ligate to unzipping construct (I have some digested pBR322 and will use this in another reaction)
 * Purify product
 * Run gel of product with neutravidin to test biotin-avidin bonding
 * Perhaps try with anti-dig as well
 * Would I need to do it on PAGE?

Digestion
http://spreadsheets.google.com/pub?key=0Agbdciapt4QZdGs2cDRrdXdDelJZWV81ZzFYWEY1MlE&output=html

Ligation
http://spreadsheets.google.com/pub?key=0Agbdciapt4QZdFAtdW9sWTRVYmlBY0ROM3dtTlZvSXc&output=html