NAME SECTION pKAN Lab (1.3 pt) 5a. (0.4 pt) In the boxes below, sequence to indi

Question

NAME SECTION pKAN Lab (1.3 pt) 5a. (0.4 pt) In the boxes below, sequence to indicate the cut site position on the complementary write the complementary sequence and draw an "up" arrow within the strand. EnzymeRECOGNITION SEQUENCE A GAT CT TACTRGA G CG CT C G A G GTC CC C G G G b. (0.2 pt) Which enzymes cut leaving blunt ends? c. (0.2 pt) Which enzymes cut leaving ends with 5' overhangs? n enzymes with different recognition sequences but which leave the d. (0.2 pt) Isocaudomers are restrnctio same sticky ends. Which two enzymes above are isocaudomers? site each for these two isocaudomers. asmid has one cof the fragments and then clone the fragment into pBluescri isocaudomens mes), would the fragment be able to insert in only one o Assume you cut the plasmid with orientations? Explain your answer

Explanation / Answer

The fragment will be able to insert in only one possible direction.

Explaination:

Lets say ATGCTAGCTAGATCTAG||AGATACGATCTAG||AGC

double dash marks the restriction site

After restriction, the sticky ends generated will be:

5-'GATC-3'

3'-CTAG-5'

Note: Pleas remember since its a plasmid it would be circular in shape, the first nick would produce a linear plasmid and the second would give two fragments. To successfully clone, the fragment should carry the compatible sticky end. Therefore on CTAG can bind with GATC therefore there can be only one orientation. The other end would carry a different sequence and would pair up with the sticky end at the pBluescript.

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