Compatibility of PCANTAB5E and M13cp-CT

2015-01-13

azim58 - Compatibility of PCANTAB5E and M13cp-CT

see Two Plasmids One E coli for more information

Compatibility of My Origins of Replication
PCANTAB5E: F1 origin of replication
M13cp-CT: based on M13mp19 plasmid which has the F1 origin of replication
-If both plasmids truly do have the F1 origin of replication, then they
are not compatible.
-I suppose I could order primers and use PCR to check for sure.
-If it does have the F1 origin of replication, I would need to replace it
with a different origin of replication somehow.
Compatibility table:
http://www.bio.davidson.edu/Courses/Molbio/Protocols/ORIs.html
>F1 Sequence
gcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcct
ttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctcccttta
gggttccgatttagtgctttacggcacctcgaccccaaaaaacttgatttgggtgatggttcacgtagtgggcca
tcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaact
ggaacaa
Length: 309
To check if my M13cp-CT and M13cp plasmids really do have this sequence I
will need to do a colony PCR.
The Primer3 website recommends these primers
Primer_1_F
F1_F
ctcctttcgctttcttccct

Start: 70
 Length: 20
 Tm: 59.955
 GC%: 50.000
 ANY: 2.00
 SELF: 2.00

Primer_1_R
F1_R
aagggcgaaaaaccgtctat

Start: 253
 Length: 20
 Tm: 59.966
 GC%: 45.000
 ANY: 3.00
 SELF: 3.00

Product Size: 183
 Pair Any: 6.00
 Pair End: 1.00

Link to order form for these primers:
http://www.wuala.com/azim58/wuala/CIM%20Research%20Folder/kwhittem/Records%
20in%20CIM%20Folder/Dated%20Records/2011/5-11-11/F1%20Ori%20Primers.xls/

Link to receipt for these primers:
http://www.wuala.com/azim58/wuala/CIM%20Research%20Folder/kwhittem/Records%
20in%20CIM%20Folder/Dated%20Records/2011/5-23-11/F1%20ori%20primer%20Receip
t.pdf/

azim58wiki: