Allelic Expression Analysis of Imprinted Genes

2.2.1. Total RNA Purification and Reverse Transcription

1. Dounce homogenizer (7 mL).

2. Isogen (Nippon Gene).

3. DNase I (RNase-free; Nippon Gene).

4. Phenol.

6. Ethanol.

7. SuperScript III reverse transcription (RT) kit (Invitrogen). 2.2.2. RT-PCR and Allelic Expression Analysis

2. DNA sequencer: 3100 Genetic Analyzer (Applied Biosystems).

3. ExoSAP-IT (Amersham Biosciences).

4. BigDye Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems).

5. Appropriate primers for DNA sequencing.

6. Thermal Cycler: GeneAmp PCR System 2400 (Applied Biosystems).

7. Appropriate restriction enzymes.

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