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Samsung

Nimble footed global adoption of the Android OS, Samsung is quick moving towards leadership position. Even in India it is the #1 Android based phone brand.

In addition it has made the “smart” positioning its very own, both at retail and ATL

“smart”

Low cost of entry into the Android OS and the brand equity of the parent brand, a trusted Consumer Goods player in India for years has benefited Samsung immensely. Last fiscal, more than ever Samsung leveraged this advantage andhas been quick to release new products at different price points, biting into significant share of the market while Nokia bided its time on what position it wants to acquire in the future. However, Nokia with its well reviewed range of Lumia Smartphones will be a threat again and 12-13 must test the endurance of Samsung as it will feel the heat from all quarters.

Rank #2 – Fastrack

Rank #2 – Fastrack

Continuing its great work from the Move On campaign (released in 10’-11′),Fastrack came out with peculiar rationales as to why the world indeed moved on .Persisting with Genilia and Kohli as their young ambassadors Fastrack connected with the young via naughty connotations to their ads.

Move On why the world moved on

These are brand ads where they demonstrate a range of their products but most importantly a personality that is vivacious.Just one critique, the hand break and the auto pilot cause duplication

Rank #1 – AIRTEL

Rank #1 – AIRTEL

Over a year back, Airtel executed a confused yet efficientre-branding exercise. This was the time it had shifted focus to its Africa ambition, one only withaChinese parallel.It was all too clear that another leader was spreading itself too thin and might break in the face of heating competition.

Until, the year past when it came up with perhaps the mostsuccessful youth campaign of the year and reestablished itself not only as the #1 telecom player but also a serious contender as the top youth brand.

Har Ek Friend Zaroori hota hai struck a chord with youth across the country and as a concept was extremely scalable and relevant. Be it pushingtheirassociation with or launching Chloé Python RoundToe Pumps Excellent Online DYie4g4
.

Overall it was a great year in advertising as it threw up some unlikely winnersreflectingthe adventure and optimism in the Indian market. These are encouraging signs as the control is with the young consumer, the ADHDed. Brands will have to take some chances and understand the youth better.

It will also call forentrepreneurial ambition in and around corporate India as new products have enough early adopters if developed and launched well. The fact that names like Vodafone, Docomo, Appy Fizz,Nokia, Micromax, Thumbs Up, Sprite, Dairly milk dont find a mention in this list is because competition is tough and youthcentricity is essential to attach a #Rank adjacent to your names.

Watch the protocol video below to learn how to isolate single bacterial colonies.

Take competent cells out of -80°C and thaw on ice (approximately 20-30 mins).

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(containing the appropriate antibiotic ) from storage at 4°C and let warm up to room temperature and then (optional) incubate in 37°C incubator.

Mix 1 - 5 μl of DNA (usually 10 pg - 100 ng) into 20-50 μL of competent cells in a microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the tube with your finger a few times.

*Pro-Tip* Transformation efficiencies will be approximately 10-fold lower for Discount Websites Stuart Weitzman Lace dOrsay Pumps Clearance Nicekicks Inexpensive Online View 5arPrD3PU
of inserts to vectors than for an intact control plasmid.

Incubate the competent cell/DNA mixture on ice for 20-30 mins.

Heat shock each transformation tube by placing the bottom 1/2 to 2/3 of the tube into a 42°C water bath for 30-60 secs (45 secs is usually ideal, but this varies depending on the competent cells you are using).

Put the tubes back on ice for 2 min.

Add 250-1,000 μl LB or SOC media (without antibiotic) to the bacteria and grow in 37°C shaking incubator for 45 min.

*Pro-Tip* This outgrowth step allows the bacteria time to generate the antibiotic resistance proteins encoded in the plasmid backbone so that they will be able to grow once plated on the antibiotic containing agar plate. This step is not critical for Ampicillin resistance but is much more important for other antibiotic resistances.

Plate some or all of the transformation onto a 10 cm LB agar plate containing the appropriate antibiotic.

*Pro-Tip* We recommend that you plate 50 μL on one plate and the rest on a second plate. This gives the best chance of getting single colonies, while allowing you to recover all transformants.
*Pro-Tip* If the culture volume is too big, gently collect the cells by centrifugation and resuspend in a smaller volume of LB so that there isn't too much liquid media on the agar plates. If the agar plate doesn't dry adequately before the cells begin dividing, the bacteria diffuse through the liquid and won't grow in colonies.

Incubate plates at 37°C overnight.

If you are not concerned with transformation efficiency (such as when you have a tube of plasmid DNA and just need to transform bacteria so that you can grow up more of the plasmid) you can save a lot of time by shortening or skipping many steps and will still get enough colonies for your next step. Remember that each of these shortcuts will reduce the efficiency of the transformation, so when higher efficiency is needed follow the complete protocol.

Chemically competent cells are fast and easy to use, but are less efficient at taking up larger plasmids. If you need to transform large plasmids, it is a good idea to use electro-competent cells. Instead of relying on the heat-shock to cause the cells to take up the DNA, an electro-magnetic field is applied to the cell/DNA mixture to induce membrane permeability. To do this you will need to have access to an electroporator and the appropriate cuvettes. Follow the manufacturer's instructions for each.

Check that you are plating on an LB Agar plate containing the correct antibiotic. The resistance gene on your plasmid must match the antibiotic on the plate. You should also add a positive control (many companies include a positive control plasmid with their competent cells) to ensure that your transformation procedure is working.

Although it may be counter-intuitive, you will often get higher transformation efficiencies with less DNA, especially when using highly competent cells. If you used 100-1000 ng of total DNA in a ligation you will often get more colonies if you use 1 μl of a 1:5 or 1:10 dilution rather than 1 μl directly.

How can I save time when carrying out transformations?
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Introduction

Transformation is the process by which foreign DNA is introduced into a cell. Transformation of bacteria with plasmids is important not only for studies in bacteria but also because bacteria are used as the means for both storing and replicating plasmids. Because of this, nearly all plasmids (even those designed for mammalian cell expression) carry both a bacterial origin of replication and an antibiotic resistance gene for use as a selectable marker in bacteria.

Scientists have made many genetic modifications to create bacterial strains that can be more easily transformed and that will help to maintain the plasmid without rearrangement of the plasmid DNA. Additionally, specific treatments have been discovered that increase the transformation efficiency and make bacteria more susceptible to either chemical or electrical based transformation, generating what are commonly referred to as 'competent cells.'

Many companies sell competent cells, which come frozen and are prepared for optimal transformation efficiencies upon thawing. For the highest transformation efficiency, we recommend that you follow the instructions that came with your competent cells.

For the highest transformation efficiency, we recommend that you follow the instructions that came with your competent cells.
*Pro-Tip* Commercial competent cells range significantly in their transformation efficiency. The lowest efficiency cells (usually the least expensive) are fine for transforming plasmid DNA for the purposes of storage and amplification. Higher efficiency cells are more important if you will be transforming with very small amounts of DNA or if you're multiple plasmids at once.
*Pro-Tip* To save money, many labs also make their own competent cells. This is a relatively simple procedure and is useful for performing low efficiency transformations.

Last Update: Nov. 13, 2017

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