User:Daniel Goodman/Notebook/Weiss/2010/01/19: Difference between revisions

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Line 32: Line 32:
| 2 || PIF3 || ~15ul || 1.5ul
| 2 || PIF3 || ~15ul || 1.5ul
|-
|-
| 3-5 || PhyB917 Samp 1-3 || ~15ul || 1.5ul
| 3 || PIF3 Old || ~15ul || 1.5ul
|-
| 4-10 || PhyB917 Samples 1-7* || ~15ul || 1.5ul
|-
|}
|}
''* Samples 1-3 are new, 4-7 are recovered from freezer from Dec, may fail''


===Expected for PhyB917:===
===Expected for PhyB917:===

Revision as of 12:05, 19 January 2010

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Restriction and Gel of PhyB917 & PIF3

Samples First 3 PhyB917 Samples (37 deg C, 10 min) PIF 3 Sample (65 deg C, 10 min)

Made 75 ml agar gel:

  • 75ml TAE
  • .75 g Agarose
  • 7.5 ul SYBR-SAFE

Put SYBR after cooling, cooled in water bath, set gel ~20 min.

Note: forgot to swirl after adding sybr, though color looked uniform

Cut top of gel off, saved bottom in 4°C fridge in ziploc w/ name&date

Loading Gel:

Lane Sample Amt Loading
1 Hyperladder 1 5 ul n/a
2 PIF3 ~15ul 1.5ul
3 PIF3 Old ~15ul 1.5ul
4-10 PhyB917 Samples 1-7* ~15ul 1.5ul

* Samples 1-3 are new, 4-7 are recovered from freezer from Dec, may fail

Expected for PhyB917:

Cut with BstBI - two bands, 3579 and 5314

Expected for PIF3:

Cut with AvrII - two bands, 2164 and 4791