Table 18.2 Characteristics of some common type II restriction enzymes used in recombinant DNA technology Microorganism from Type of fragment Enzyme which enzyme is isolated Recognition sequence end produced BamHI Bacillus amyloliquefaciens 5-GGATCC-3 Cohesive 3'-CCTAGG-3' EcoRI Escherichia coli 5'-GAATTC-3 Cohesive 3'-CTTAAG-5' EcoRII Escherichia coli 5-CCAGG-3' 3'-GGTCC-5' Cohesive Haelll Haemophilus aegyptius 5-GGCC-3' 3-CCGG-5 Blunt HindIII Haemophilus influenzae 5'-AAGCTT-3' 3'-TTCGAA-5 Cohesive Notl Nocardia otitidis-caviarum 5-GCGGCCGC-3 Cohesive 3'-CGCCGGCG-5 Pstl Providencia stuartii 5-CTGCAG-3' Cohesive 3'-GACGTC-5' ↑ Smal Serratia marcescens 5-CCCGGG-3' Blunt 3'-GGGCCC-5'

Create the restriction map for the plasmid below:

A cloning vector map is shown below. EcoRI Bam Ban Hind P-galactosidase Amp Bam Bam EcoRI Ori C Which restriction site is best for inserting a DNA fragment for selection of chimeric plasmid containing colonies? 1) They're all equally good. 2) Hindll 3) EcoRI 4) BamHI

Use the gel to answer the following questions. You will be constructing a map of the plasmid, pDiddy.   What is the smallest fragment size that the NcoI/EcoRI double digest produces?

Use the gel to answer the following questions. You will be constructing a map of the plasmid, pDiddy.   IF the EcoRI/BamHI double digest produces 3 fragments with only two sizes, what are their sizes?

After cloning, they transformed and plate bacterial cells using their cloned plasmid. Onto what type of growth medium will they plate their cells in order to distinguish between bacterial cells that obtained the plasmid and those that did not?

The DNA polymerase used in PCR is taken from Thermus aquaticus, a bacterium that thrives in the extremely high temperatures of hot springs. Why is it necessary to use special DNA polymerase? * bers erences QOnline Thermus aquaticus is accustomed to efficient replication. wsela Thermus aquaticus can survive at boiling and freezing temperatures, both of which are necessary during PCR. ation ogy Periods 1 and 2 "Normal" DNA polymerase would denature in the high temperatures used in PCR. ng periods chool MP1, Highschool Highschool MP3, school MP4 Thermus aquaticus is able to supply a varied amount of bases to the DNA template. tion ting days n Tue Wed Thu Fri Mutations occur randomly during the process of gene expression or by exposure to mutagens. While some mutations are detrimental, others can create proteins that ha n nociti e conahilition In vder fo ra mutation te be n esed tn futuen INTL

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Use the gel to answer the following questions. You will be constructing a map of the plasmid, pDiddy.   What is the largest fragment size that the BamHI/NcoI double digest produces?     2.250 kb     2.500 kb     2kb     750 b

In Figure 5-19, how many different bacterial species areshown as having contributed DNA to the plasmid pk214?