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& Routine Cloning Phage Display Library Construction Toxic/Unstable Dna Cloning High-Throughput Cloning Browse Full Report with TOC: https://www.
We used the reverse transcription PCR method and subcloning
protocol (online Technical Appendix, https://www.
On the base of recent study experiences, the chance of getting stable hybridoma with high mAb titer was dependent on cell culture medium, appropriate screening and early subcloning
methods in aseptic conditions.
Now an encouraging study by scientists at the NIH's National Human Genome Research Institute (NHGRI) suggests that iPSCs do not develop more mutations than cells that are duplicated by subcloning
, a technique in which single cells are cultured individually and then grown into a cell line.
To localize the AY-glucosidase gene and shorten the insert for effective analysis of enzymology characterization, subcloning
USA) was used for subcloning
and yeast vector amplification.
of the best CHOBRI/Cum2 pools, generated by transfection, increased productivity by 3 to 6-fold yielding clones producing up to 1.
ipaB in pBAD/gIII expression vector and its transformation into E.
Leachman for practical advice concerning RT-PCR and subcloning
The amplified product was purified by PCR purification kit following manufacturer's instructions (Qiagen, Germany) and cloned first in the TA cloning vector pTZ57R (Fermantas) followed by subcloning
in the yeast shuttle expression vector pGal426 that contains the galactose inducible promoter Gall and uracil synthesizing gene (ura3) as selection marker (12,13).
We then submitted these sequences to Blue Sky Biotech for subcloning
and in vitro transcription.
The Morex wild-type rBmy2 enzyme had 16 extra amino acids attached to its N-terminal end, artifacts from subcloning
into the expression vector, while the Steptoe wild-type rBmy2 had six extra amino acids on its N-terminal end.