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

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

Things I've Tried

• washed invitrogen fluospheres to remove streptavidin, resuspend in BGB
• concetrated anti-dig
• incubate beads with DNA
• high salt beads during tethering, wash with BGB

Brian is running the PCR to make more 4.4kb stretch DNA.

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