For clones lacking an oriV this sequence can be easily retrofitted into the clone either by recombineering or by standard genetic engineering techniques or, and perhaps more straightforwardly, via random Tn5 transposon-mediated insertion [ 18] (EZ-Tn5 system, Epicentre).
Transgenic animals were synchronised and subject to the acute RNAi protocol with scoring occurring at 24 hours post introduction to RNAi clone or a control clone lacking a targeted sequence.
We did not sequence a DNA clone lacking an insert, but the mitochondrial genome of other dinoflagellates is known to contain many copies of each gene and many rearrangements [ 21, 23], so the intact copy of the gene was most likely simply not sampled.
In the 2-hour series finale, it is revealed Lex had used the parts of other clones to create a perfected one; however, the clone lacked a heart.
The U5A clone lacks a functional IFNAR2 receptor (IFNAR2−/−) and thus is impaired in type I interferon receptor signaling (Lutfalla et al., 1995).
When we used an e-value of >0.1 to identify such clones, we found that 3.1% of the BBRAF clones lacked any Arabidopsis homologues (Fig. 1B).
The elegance of the GalK based recombineering system [55] relies on the ability to counterselect for clones lacking GalK using deoxygalactose, which is metabolised to a toxic intermediate.
pELMO was thus selected as an alternative choice for simplifying cloning and avoiding the occurrence of clones lacking inserts; this newly designed in-house vector is a versatile efficient tool which is suitable for cloning blunt-ended PCR-products as it relies on the effect of a toxic gene from the pDONR221 sequence.
Similar to results in bulk cells, clones lacking the upstream promoter region of MALAT1 do not display a strong reduction in RNA levels (Fig. 6a).
Consequently, the observed number of clones lacking TdCRT1 hybridization is 642 times (16,751/26 = 642) the expected standard deviation for a normal distribution, strongly supporting the premise that the distribution of TdCRT1 is not random.
