Of that 14-week development time,
subcloning and optimal clone identification still typically require four to eight weeks.
This
subcloning was further confirmed by subsequent restriction digestion analysis (Figure 1).
pGEX-MED6 was made by
subcloning the EcoRI-SalI fragment from pGBT-MED6 into pGEX-2TK (Pharmacia).
After the first cycle of
subcloning, five clones stably produced specific antibodies, while the other three clones were determined to be negative (Table 1).
Subcloning of Lipase Genes From Cloning to Expression Vector
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.
coli DH10B and the vectors pUC18 and pCR2.1 (Invitrogen Life Technologies, USA) were used for
subcloning steps.
To localize the AY-glucosidase gene and shorten the insert for effective analysis of enzymology characterization,
subcloning was performed.