IGEM:IMPERIAL/2006/project/Oscillator/project browser/Test Killing Predator Construct/Implementation: Difference between revisions
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**B0014 (1G) (vector) cut with EcoRI & XbaI, expected size = 95 bp + 3189 bp (pSB1AK3) = 3284 bp | **B0014 (1G) (vector) cut with EcoRI & XbaI, expected size = 95 bp + 3189 bp (pSB1AK3) = 3284 bp | ||
*Miniprep sizes expected: 911 bp & 3189 bp (pSB1AK3) | *Miniprep sizes expected: 911 bp & 3189 bp (pSB1AK3) | ||
Revision as of 20:24, 29 October 2006
Super Parts | Predator Construct | |||
---|---|---|---|---|
Actual Part | AiiA Testing Construct <bbpart>J37022</bbpart> | |||
Sub Parts | <bbpart>R0010</bbpart> | <bbpart>B0034</bbpart> | <bbpart>J37023</bbpart> | <bbpart>B0015</bbpart> |
Ligation Process
Step 1: Construct <bbpart>J37023</bbpart> from Part <bbpart>C0060</bbpart> and a FLAG immunotag by PCR (sequence from Dr. Mann)
Step 2: Construct <bbpart>J37024</bbpart> by adding a terminator (<bbpart>B0015</bbpart> to J37023
Step 3: Construct <bbpart>J37022</bbpart> by adding the LacI promoter (<bbpart>R0010</bbpart>) and the RBS <bbpart>B0034</bbpart>, which is contained within the part <bbpart>J04500</bbpart>.
People Involved with Ligations:
- John Sy
- ???
<bbpart>J37022</bbpart> (Final AiiA test construct)
- <bbpart>J04500</bbpart> (16P) + <bbpart>J37024</bbpart>
- J37024 (insert) cut with XbaI & PstI, expected size = 911 bp (on pSB1AK3 = 3189 bp)
- J04500 (16P) (vector) cut with SpeI & PstI, expected size = 220 bp + 3189 (pSB1AK3) = 3409 bp
- Miniprep sizes expected: 1131 bp & 3189 bp (pSB1AK3)
<bbpart>J37023</bbpart> (for predator cell/test construct)
- AiiA with Immunotag
- Done by PCR
- Status: Complete (awaiting sequencing results)
- Expected Part Length = 816 bp
- Expected Vector Length = Unknown (Hanging T tail vector)
<bbpart>J37024</bbpart> (for AiiA predator cell/test construct)
- <bbpart>J37023</bbpart> + <bbpart>B0015</bbpart>
- J37023 (insert) cut with EcoRI & SpeI, expected size = 816 bp
- B0014 (1G) (vector) cut with EcoRI & XbaI, expected size = 95 bp + 3189 bp (pSB1AK3) = 3284 bp
- Miniprep sizes expected: 911 bp & 3189 bp (pSB1AK3)