Already have an account? Get multiple benefits of using own account!
Login in your account..!
Remember me
Don't have an account? Create your account in less than a minutes,
Forgot password? how can I recover my password now!
Enter right registered email to receive password!
Sally Subcloner wants to clone a fragment into pUC19 (Map above - From New England Biolabs). Her fragment was a protein coding gene of 2.7 kb, and had XhoI sites on either end (and no other XhoI sites any where else) and she agarose gel purified this fragment to ensure it was pure. As no XhoI sites existed in the pUC19 plasmid, she decided to clone it into the Sal1 site, as Sal1 and Xho1 have identical overhangs of 5'-TCGA-3' and therefore she believes that they can be ligated to one another. As she was using blue/white screening she did not bother with Alkaline Phosphate treatment and ligated the XhoI fragment directly with SalI digested pUC19. After transformation into E. coli (strain AG1 using heat shock treatment) she plated cells onto media with Ampicillin selection (100 mg/mL) and obtained over 300 ampicillin resistant colonies. However none of these were white colonies, only blue, which she interpreted as obtaining no plasmids with the fragment she was aiming to subclone.
(a) Was her cloning approach valid and what was the reason that she could not get white colonies from this protocol? If she modifies the protocol to address this question, what additional advice can you give Sally Subcloner to increase her chances of obtaining white colonies.
(b) After taking this advice and obtaining white colonies, she purified the plasmid from five different white colonies digested them with Xho1 and ran them on a gel (see below). However there were three bands in each lane, with the major band running at approximately 4.5 kb, instead of 2.7 kb that she expected her XhoI fragment to be. Could these plasmids still be the right recombinant and what simple experiment could she perform to determine this? (Hint what control should have been included in the gel?)
What is meant when it is said that a virus is in an inactive state? Viruses considered in inactive state are those whose genetic material is within host cells without synthesis
Q. What is low - density lipoprotein receptor pathway? Ans. The increasing cellular free cholesterol generated regulates the activities of two enzymes that are of crucial
Define Descending type of paper chromatography? In descending chromatography, the movement or flow of mobile phase is in downward direction. In this technique the solvent is ke
Explain Transfer from Solid Culture to Liquid Media The steps involved in this technique are included herewith: 1. Sterilized inoculating loop or needle is touched carefully
Ethylene - Plant Hormones Amongst hormones in both plant and animal kingdoms, ethylene, a gaseous hydrocarbon, is unique. Despite its chemical simplicity, it is a potent growt
Q. What is the difference between cellular specific immune response and humoral specific immune response? Humoral specific immune response is the defense system by means of, an
BRANCHES OF EMBRYOLOGY- 1 . DESCRIPTIVE EMBRYOLOGY This field of embryology is associated with the morphological description of different embryonic stages in the on
Feline panleukopenia It is arguably the most important of all feline viral diseases and all members of the family Felidae are susceptible. This disease is commonly found in kitten
The ability to sequence DNA has led to a revolution in molecular biology. a) Describe the principles of DNA sequencing. b) A single sequencing reaction yields at best 1500 ba
Consider Neuron B in the frog central nervous system whose plasma membrane has a previously unknown channel that is selectively conductive to a newly discovered tetravalent anion n
Get guaranteed satisfaction & time on delivery in every assignment order you paid with us! We ensure premium quality solution document along with free turntin report!
whatsapp: +91-977-207-8620
Phone: +91-977-207-8620
Email: [email protected]
All rights reserved! Copyrights ©2019-2020 ExpertsMind IT Educational Pvt Ltd