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?)
Q. Causes of gastro oesophageal reflux disease? GERD may develop due to any of the following reasons: • decreased muscle tone or abnormal relaxation of the LES, • reduced st
Write up your own Matlab program to do the job described by Show">http://www.avatar.se/molbioinfo2001/dynprog/adv_dynamic.html Show a few examples, your code does work. You wil
Most proteins made by ribosomes on the RER (rough endoplasmic reticulum) are glycoproteins, which is they hold short chains of carbohydrates (oligosaccharides) covalently related t
Advantages of Metagenesis in Cnidaria In this you have studied that a huge number of Cnidarian colonies are polymorphic and exhibit alternation of generations. This changes o
why protozoass dont have vaccines esp. plasmodiums
Q. Definition of Consciousness? Consciousness is a phenomenon that is shared by nearly all people. It cannot be directly seen or touched. Yet it is real enough to most people.
How cholera is generally spread in humans Associated Foods: Cholera is generally a disease spread by poor sanitation, resulting in contaminated water supplies. Sporadic cases o
Pre Pregnancy Counselling To prevent excess spontaneous abortions and congenital malformations in infants of diabetic mothers, diabetic care, education and counselling must be
endoplasmic reticulum
SCOPE OF BIOLOGY- Present century is witnessing explosion of information in modem biology directly related to life and health. Issues like global warming, pollution, populat
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