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Engineering PC-TFs
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Summary
Cutting inserts to be placed in mammalian vector
A) Get biobricks and enzymes from freezer.
SpeI, PstI, XbaI
The following list of parts are:
Stock Digest
B)Follow digest procedure:
DNA 20.0 µL
Enzyme 1 1.0 µL
Enzyme 2 1.0 µL
10x buffer 3.0 µL
dH2O 5.0 uL
= 30.0 uL
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Digest Order:
Biobrick Restriction Enzymes
- 1. BL01 XbaI/PstI
- 2. BL05 XbaI/PstI
- 3. BL09 XbaI/PstI
- 4 BL12 XbaI/PstI
Reagent
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Volume
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30 μL/lane, 1% agarose; Ladder
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DNA (plasmid) |
20.0 μL
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10x buffer |
3.0
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XbaI |
2.0
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dH2O |
5.0
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30 μL --> 37°C/ ~15 min.
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#
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Biobrick
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ng/µL
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1
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hPCD : mCh : SP1AB (BL01)
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18.834
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2
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fshPCD : mCh : SP1AB (BL05)
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14.471
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3
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flyPCD : mCh : SP1A (BL09)
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7.302
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4
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hPCD : BL01 (BL12)
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10.84
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