Of that 14-week development time, subcloning
and optimal clone identification still typically require four to eight weeks.
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).
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
the cells of clone 3G5, the results were similar to those from the initial cloning.
and Detection of Overexpression and Deacetylase Activity of HDAC6.
To localize the AY-glucosidase gene and shorten the insert for effective analysis of enzymology characterization, subcloning