Team:Kyoto/GSDD/Notebook/0907-0910



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To Do
(8) (7)(8)(10)(5)
 * Make repetitive sequence (MPR)
 * Make parts(for flush end ligation)
 * Annealing
 * Restriction Enzyme Digestion
 * Ethanol precipitation
 * Ligation
 * transformation

MPR

 * Electrophoresis results below indicated that sample E and F are proper for extracting.

Make parts(for flush end ligation)

 * This indicated that annealing product was not obtained.

Transformation

 * (5)A,(5)B:Colonies were observed.
 * (7)A,(7)B,(8),(10)A,(10)B:No colony was observed.

To Do

 * MPR products (further to 7 Sept.)
 * Gel extraction
 * Measure concentration
 * Make repetitive sequence (MPR)
 * (1)(4)(5)
 * Colony PCR
 * Electrophoresis
 * Miniprep and make master plates

Make repetitive sequence (MPR)

 * Gel extraction and Measured concentration.

Colony PCR of (1),(4),(5)

 * This result indicates that each part were inserted properly.

To Do

 * PCR products(1)(4)(5)(0908)
 * Gel extraction
 * Measure concentration (See lab note on 8 Sept.)
 * PCR product(5)
 * Miniprep and make master plates
 * (7)(8)(10)
 * Ligation
 * Transformation
 * Make parts(for flush end ligation) retry
 * Annealing
 * Restriction Enzyme Digestion
 * Ethanol precipitation

(7),(8),(10)
We failed in the transformation 0907 because of the plasmid we used. We tried again using plasmid we purified anew.
 * PSB1A2(ES) conc.: 24(ng/ul)

After transformation, colonies were observed in all plates.

To Do

 * (7)(8)(10)
 * Colony PCR
 * Electrophoresis
 * PCR

(7)(8)(10)
Inserted properly This result indicates that some samples were inserted properly, though others were self ligated.
 * (7)A①③,B②
 * (8)①
 * (10)A①②③,B②③

PCR
This indicates PCR was achieved.