Serial Analysis of Gene Expression Using ABI PRISM DNA Sequencers and BigDye Terminators
Serial Analysis of Gene Expression (SAGE) is a sequence-based approach to the identification of differentially expressed genes by comparative analyses. SAGE is a powerful tool, one of the more comprehensive methods available for detailed analysis of large numbers of cellular transcripts, ...
Comparison of Commercial Dye Terminator Removal Kits, Rev A
...cturers were assessed using these four criteria (Table 1).
Cycle sequencing was performed using bigdye
terminators, version 3.0 (Applied
Biosystems). The plasmid template used was pGEM-3Zf(+), acquired from
Promega. The primer used...
Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
...en was used
for cloning. The agarose gel extraction kit and plasmid extraction kit
were purchased from QIAGEN Ltd. The ABI PRISM bigdye
terminator v 3.0
cycle sequencing kit with AmpliTaq DNA polymerase, FS was purchased from
Applied Biosystems. The TaqMan reagent...
cDNA Sequencing & Finishing
Data delivery via a secure FTP site
SEQUENCING AND FINISHING
Purified cDNA clones are sequenced using bigdye
Terminator v3.1 chemistry
and reactions purified using CleanSEQ, a SPRI-based dye-terminator removal
system. Typical Phred20 rea...
CleanSEQ on the ABI 3100
pGEM was sequenced using v3.1 bigdye
chemistry at 1/4x dilution. Sequencing reactions wer...
CleanSEQ with water elution allow extensive
dilution of bigdye
Terminators without affecting
Phred20 read lengths (data not show...
SPRINTPREP SINGLE STEP PLASMID PURIFICATION
...00 cDNA clones as part
of the Mammalian Gene Collection. Purified plasmids were sequenced using
30ng of template DNA in a 1/16th bigdye
v3.1 terminator reaction. Unincorporated
dye-terminators were removed using CleanSEQ and sequences read on ABI
3700 and 3730xl's...
DirectPrep 96 Miniprep System for cost-effective, high-throughput plasmid
sequencing (Figure 3). Using either bigdye
terminator v.2.0 or v.3.0
sequencing chem... and
reactions were sequenced using bigdye
terminator v. 2.0 chemistry
and ana... Kit. Plasmid pUC19 was sequenced using bigdye
terminator v. 3.0 chemistry
in a cy...