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Due to this variability of sarcomere length during contractions, all results presented herein are related to sarcomere length readings made prior to the contractions (passive sarcomere length), at one position along the length of the fiber.
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Today's full-length readings of the two indictments, an unusual event here and a procedure that is ordinarily waived, was the court's response to the ongoing defiance of Mr. Milosevic, who continues to call the court illegal and has scorned all the court documents that have been delivered to him in his United Nations prison cell here.
Assembly of full length open-reading frames for three additional genes was done accordingly.
cDNAs encoding full-length open-reading frames (ORFs) of BmRas1, BmRas2 and BmRas3 were amplified by PCR using primer pairs ras1-NdeIF and -BamHIR, ras2-NdeIF and -BamHIR and ras3-NdeIF and -BamHIR, respectively (Table 1), and ligated into pET-16b plasmid vector (Merck KGaA, Darmstadt, Germany).
With the exception of one FPS, the full-length open-reading frames (ORFs) of other unigenes were unavailable.
The pcDNA-DEST53 plasmid containing GFP fused to the N-terminus of the full-length open-reading frame (ORF) of human PERP (GFP PERP) under the CMV promoter was constructed as described previously.
The full-length open-reading frames of RACK1A (At1g18080), RACK1B and RACK1C were amplified from a cDNA library made from seedlings grown in light for 10 d, cloned into the pENTR/D-TOPO vector (Invitrogen, ), and then subcloned into Gateway plant transformation destination binary vector pB2GW7 [ 22] by LR recombination reactions.
To generate HA or GD tagged constructs for TCL2, SPL3, 9, 10, 13 and 15, the full-length open-reading frames (ORF) of corresponding genes were amplified by RT-PCR using RNA from 10-day-old light-grown Arabidopsis seedlings, then the PCR products were cloned in frame with an N-terminal HA or GD tag into the pUC19 vector under the control of the double 35S enhancer promoter of CaMV.
Constructs used for protoplasts transfection were generated by first amplifying the full-length open-reading frame (ORF) of the corresponding genes by RT-PCR using RNA isolated from 10-d old, light-grown Arabidopsis seedlings, then cloning the PCR fragment in frame with an amino terminal HA or GD tag into the pUC19 vector under the control of the double 35S enhancer promoter of CaMV [ 36, 39].
This is due to an increase in the refractive power of the lens, since axial length and keratometry readings do not reveal a corneal or length basis for a myopic shift.
Although these tools generally provide good results, qualitative validation is important (e.g. by assessing the length of open-reading frames).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com