User:Anugraha Raman/Notebook/iGEM 2010/2010/06/25: Difference between revisions
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**Total: 40 uL Volume | **Total: 40 uL Volume | ||
**Normal Digest (Digested for 1 hour) | **Normal Digest (Digested for 1 hour) | ||
*PCR amiRNA | |||
**1 uL of 164.8 ng/uL DNA template (RS300) | |||
**.5 uL Phusion Polymerase | |||
**10uL 5x Phusion Buffer | |||
**1uL 10nm DNTP | |||
**2.5 uL fwd+rev primer | |||
**35 uL H20 | |||
**Total Volume 50uL reaction | |||
*3 Reactions | |||
**Reaction 1: Fwd Primer (A) Rev Primer ("primer 4") for LTP/GFP/Betv1 | |||
**Reaction 2: Fwd Primer "primer 3" Rev Primer "primer 2" for LTP/GFP/Betv1 | |||
**Reaction 3: Fwd Primer "primer 1" Rev Primer (B) for LTP/GFP/Betv1 | |||
**Annealing Temp for the three GFP reactions was 67 C and annealing temp for rest of reactions was 60 | |||
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