By Shel Silverstein
From Shel Silverstein, the New York Times bestselling author of Where the Sidewalk Ends and The Giving Tree, comes a riotous rhyming photograph booklet a couple of boy and his giraffe!
Featuring rhythmic verse and iconic illustrations, A Giraffe and a Half will go away each reader, old and young, guffawing until eventually the very finish. cherished for over fifty years, this vintage captures Silverstein’s signature humor and style.
If you had a giraffe and he stretched one other part, you'll have a giraffe and a half. yet what occurs in the event you glue a rose to the end of his nostril? Or in case you used a chair to sweep his hair? sign up for this giraffe on a rollicking and ridiculous trip that might allure readers from starting to finish.
Read or Download A Giraffe and A Half PDF
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Extra resources for A Giraffe and A Half
Use the following pair of primers to amplify a DNA fragment that contains a FLAG-encoding fragment followed by a cmR gene that is flanked two AscI cut sites. 1. Flag-AscI-Cm: gactacaaagacgatgacgacaagGGCGCGCCagccagtatacactccgcta Sequence in lower case encodes FLAG. The AscI site is in upper case. The italicized sequence in lower case is homologous to cmR. 2. AscI-Cm: GGCGCGCC ctgtggaacacctacatctg The AscI site is in upper case. The italicized sequence in lower case is homologous to cmR. 2. Clone the above PCR product using the TOPO cloning kit from Invitrogen according to manufacturer’s instruction.
We choose to add two adenosines immediately upstream of the BstEII cut site in the reverse primer. This puts the coding sequence in frame and creates Genome Manipulations with Bacterial Recombineering… 21 our desired mutation. The sequence AAGGTCACC when translated in the reverse orientation encodes Gly Asp Leu. Step 2. 3, insert this cmR PCR product into the master clone. Use BstEII enzyme to excise the cmR gene. 7 is limited by the number of suitable restriction enzymes. This limitation is placed not only on the residues that can be mutated, also on the exact amino acid to which a particular residue can be mutated.
4 N NaOH. 7. Put a gel upside down atop of the platform (so as DNA near the bottom of the gel locates near the membrane), place a 3MM filter paper, a stuck of paper towel, flat plate and weight in this order. Multiple Genetic Manipulations in DT40 31 8. Take out a membrane, label the sides and rinse with 2× SSC buffer twice. 9. Sandwich a membrane with clean filter papers and bake at 80 °C for 2 h. 6 Southern Blotting 1. Prepare probe DNA either by PCR or excision from the plasmids and column purification.