Protease and Proteinase Trials: Difference between revisions
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2 columns/strips of sorted chromosomes were collected from Sam. The strips were separated as follows: | 2 columns/strips of sorted chromosomes were collected from Sam. The strips were separated as follows: | ||
| H | | | H | - Head | ||
Row 1 | o | - No chromosomes | Row 1 | o | - No chromosomes | ||
Row 2 | o | - 1 chromosome | Row 2 | o | - 1 chromosome | ||
Line 23: | Line 23: | ||
Row 8 | o | - 23 chromosomes | Row 8 | o | - 23 chromosomes | ||
| A | - Tail | | A | - Tail | ||
Protease was added as follows: | |||
Rows 1,2,5,8: No P. added | |||
Rows 3,6: 1uL P. added | |||
Rows 4,7: 0.5uL P. added | |||
The same was applied to both strips, labelled Strip 1 and Strip 2 | |||
===Incubation=== | |||
Strip 1: 37C, 10min -> 70C, 15min | |||
Strip 2: 37C, 20min -> 70C, 15min |
Revision as of 22:32, 7 April 2010
Two different denaturants were used:
- Protease (Qiagen) - abbreviated P.
- Proteinase K (Invitrogen) - abbreviated P.K.
Solution Preparations
The protease (Qiagen) was easily reconstituted: 7uL water per vial
See Proteinase K Protocol for preparation of Proteinase K (Invitrogen)
3/31/10
2 columns/strips of sorted chromosomes were collected from Sam. The strips were separated as follows:
| H | - Head Row 1 | o | - No chromosomes Row 2 | o | - 1 chromosome Row 3 | o | - 12 chromosomes Row 4 | o | - 12 chromosomes Row 5 | o | - 12 chromosomes Row 6 | o | - 23 chromosomes Row 7 | o | - 23 chromosomes Row 8 | o | - 23 chromosomes | A | - Tail
Protease was added as follows:
Rows 1,2,5,8: No P. added Rows 3,6: 1uL P. added Rows 4,7: 0.5uL P. added
The same was applied to both strips, labelled Strip 1 and Strip 2
Incubation
Strip 1: 37C, 10min -> 70C, 15min Strip 2: 37C, 20min -> 70C, 15min