siRNA Design Guidelines
...the 3' end of the oligonucleotide. In addition, for
cloning into the pSilencer 1.0-U6 vector, nucleotide overhangs
to the ecor
I and Apa I restriction sites are added
to the 5' and 3' end of the DNA oligonucleotides, respectively (Figure
2). In con...
New Fusion Trans-Activator Plasmids for Studying Signal Transduction
208 to 313 of rat c-Fos 12 into the BamH I and ecor
I sites of pFA-CMV. Our results ( figure
3A ) in tran...idues 1-96 of human ATF2 14,15 was inserted
into the BamH I and ecor
I sites of the pFA-CMV vector.
Consistent with previous reports,...
New Competent Cells for Highest Transformation Efficiencies
...nd the potential size bias, two plasmid vectors were used.
Stratagenes pcmv-script vector (4.2 kb) was digested
with Xho I and ecor
I; Stratagenes two-hybrid pAD-GAL4
vector (7.2 kb) was also digested with the same two enzymes. A mouse brain
cDNA library was p...
Easy Cosmid DNA Purification with StrataPrep Kits
The resulting cosmid DNA was digested to completion with ecor
...smid DNA is completely digested with
restriction enzymes such as ecor
I ( Figure
2 ). 1
Retroviral Vectors for Gene Delivery
Coding sequence for the enhanced green fluorescent protein (GFP) 9
was inserted between the ecor
I and BamH I sites in pFB
and pFB-Neo. The 293 cell-based amphotropic retroviral producer cell line
HW293-A (unpublished data) w...
Generate High-Quality, Directional Plasmid cDNA Libraries
...using Pfu DNA polymerase, 4
ligated to ecor
I adaptors and digested with Xho I. Since
on its 3 end and an ecor
overhang on the 5 end. A...4-2.1 vector predigested with Xho I and
I (XR) restriction enzymes.
Constructing Directional cDNA Libraries from Limited Amounts of RNA
sequence. After standard second-strand synthesis, a double-stranded adaptor,
the LIC-L-adaptor, which encodes the ecor
I cohesive end and an
LIC(L) sequence, is ligated to the double-stranded cDNA ends. The adaptor-ligated
cDNA is then used as a t...
Versatile Reporter Vectors for Monitoring Viral Transduction
... seen for the humanized, red-shifted A. victoria GFP (EGFP) (data
not shown). The coding sequence for hrGFP was inserted between the ecor
I and Xho I sites in the pFB vector to produce the pFB-hrGFP vector
1A). The vector was then tested in retrov...
A New Lambda Vector for Mammalian Expression
...he Lambda ZAP-CMV
vector predigested with either ecor
I only or ecor
I and Xho
I, host strains, and helper phage. In ...e. We constructed a cDNA library in
I and Xho I sites of the Lambda ZAP-CMV vector
Functional Cloning Using
ViraPort Retroviral cDNA Expression Libraries
cDNA was inserted between the ecor
I and Xho I sites of
the plasmid pFB ( ...assess the quality of both the phosphatase-treated ecor
I-digested vector as well as the ...zed. All of the
100 clones examined had ecor
I/Xho I-excisable inserts,
with an aver...
A Versatile Power Supply for All Electrophoresis Applications
3 , the SmartPower 4000 power supply was used for horizontal gel electrophoresis.
The pBluescript vector was digested with ecor
serially diluted. These samples were loaded onto a 1% agarose gel and
electrophoresed for 1.5 hours at 100 volts in 1X TAE...