NYMU Taipei/Benchman

From 2007.igem.org

Revision as of 14:50, 24 October 2007

OD and GEL checks

Notes

• A260/A280 should be around 1.70
• A260/A230 should be less than 3.00
• color key
  • problemistic (read)
  • finished (yellow)
  • await (blue)
• digestion and buffer
  • SepI and PstI (SP) digestion using buffer 2, creates back vector
  • XbaI and PstI (XP) digestion using buffer 3, creates back insert
• Assembly process
  • STEP 1. prepare insert and vector by digestion
  • STEP 2. check digestion by GEL separation
    • if band is clear at correct position (length), cut the gel and purify/extract it
    • else go back to STEP 1.
  • STEP 3. ligate insert with vector, transform it into competent cell, and extract plasmid from liquid culture
  • STEP 4. check ligation by VF2, VR PCR with GEL separation
    • if band is clear at correct position (length), cut the gel and purify/extract it (ligation is done)
    • else go back to STEP 3.

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