SBB10Ntbk-Paulina Tran: Difference between revisions
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fk0003 (R)Base Substitution FokI cggcgccattgcAattggttttatgg | fk0003 (R)Base Substitution FokI cggcgccattgcAattggttttatgg | ||
fk0002 Reverse BamHI for FokI gctagGGATCCaaaattgatctccccattgttg | fk0002 Reverse BamHI for FokI gctagGGATCCaaaattgatctccccattgttg | ||
<pre | <?pre> | ||
===Construction file for sbb08=== | ===Construction file for sbb08=== | ||
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tn0002R Reverse BglII oligo for Tn5 EIPCR | tn0002R Reverse BglII oligo for Tn5 EIPCR | ||
CCAATAGATCTcatgaattcCACTTCAG | CCAATAGATCTcatgaattcCACTTCAG | ||
<pre | </pre> | ||
Revision as of 02:48, 9 February 2010
Paulina Tran 02:00, 9 February 2010 (EST)
Construction file for sbb21
Construction of FokI+ cleavage domain basic part sbb21
PCR fk0001/fk0003 on FokI (505 bp, gp = A)
PCR fk0004/fk0002 on FokI (141 bp, gp = B)
---------------------------------
PCR fk0001/fk0002 on A+B (620 bp, EcoRI/BamHI)
Digest pBjk2741-Bca1144 (EcoRI/BamHI, 2170+910, L)
Product is pBjk2741-ssb21 {<FokI+>}
---------------------------------
fk0001 Forward EcoRI and BglII for FokI ccaaaGAATTCgtccAGATCTCAGCTGGTTaaatctgaactggaggag
fk0004 (F)Base Substitution FokI ccataaaaccaatTgcaatggcgccg
fk0003 (R)Base Substitution FokI cggcgccattgcAattggttttatgg
fk0002 Reverse BamHI for FokI gctagGGATCCaaaattgatctccccattgttg
<?pre>
===Construction file for sbb08===
<pre>
EIPCR tn0001/tn0002R on pBjk2741-Bca1144 (2230 bp, BglII)
Product is pBca9145-Btn0001 {Tn5 5’TR}
-------
tn0001 EIPCR construction of Tn5 part
CCATAagatctGTCGACTGTCTCTTATACACATCTCAACCggatcctaaCTCGCTCCTCaggcttc
tn0002R Reverse BglII oligo for Tn5 EIPCR
CCAATAGATCTcatgaattcCACTTCAG
