A Creation called Love.

Begging some amusement, from the stagnant time, I decided to have a stroll past the Riverside (from Babughat to Princep Ghat, Kolkata). It was April 01, 2012 and I was not fooling with my wish to do so. As it is, my tuitions have dried up; after 9 years of service in the arena, I was free in a Sunday evening. It was feeling good, but how long will this be, I doubt, being the workaholic mentality I harvest within me.

I assembled my Nikon babe, cleaned the face and was click-ready. Boarding the 30C/1 from Taltala stop, I reached Babughat within 15 minutes, which generally takes 25 minutes on a normal Kolkata weekday. On road, I met Eden Gardens; decorating her attire to give birth of the several great matches, to be held from 5th April, 2012 under the big banner of IPLt20! It was shaded purple-gold, being the host ground for Kolkata Knight Riders. I knew I will visit her soon, so I rode past her towards my destination.

Unknowingly, my luck favored my day. There was sounds of cymbals in the air and drums banging; it was not any puja's terminal day- or was it? It was. Goddess Annapurna was being immersed in the ganges, to complete the last rites of a night long festivities. I was lucky, to take some snaps of immersion, as I always wanted to do so. Ma Annapurna, fulfilled my wish.

Immersion, Babughat | Kolkata

(c) Somnath Paul, 2012.

Till now, my love for the camera expressed herself in proper attire; but then, the passing waters slowly drizzled down my soul. Asking your presence. It seemed that eons ago, I have had walked through the trail, under the trees, making life smile beside the flowing water. It was ever steady in her flow and so was I with my life. Then, incidents for recent past lured me to believe the lights of Vidyasagar Setu; shinning bright and shedding its ray of hopes to every tiny soul, using it to cross over the water hurdle.

Vidyasagar Setu | Kolkata

(c) Somnath Paul, 2012

Its a hurdle, that I have to cross, to reach you on the other hemisphere of this earth. Its a creation and Kolkata, remains the epicenter of all creations.

Whole Blood DNA Isolation Protocol

I am writting down the protocol for the DNA Isolation via Salting out process from blood. Gives good yield of DNA. A260/280= 1.6-1.8 (best)

Reagents

1) Solution A Haemolysis Solution * Prepare fresh or use a solution prepared in 24 hrs and stored at 4'C.

Final Volume (ml):          1000                    750                     500                  100

Sucrose (g):                   109.5                   82.125               54.75                10.95

1M MgCl2 (ml):                5.0                      3.75                   2.5                     0.5

Dissolve the above in autoclaved water. Do not autoclave after dissolution as it can char the sucrose.

Add Triton X @ 1% of the final volume.

2)  Solution B  WBC Lysis buffer * Prepare fresh or use a solution prepared in 24 hrs and stored at 4'C. Add SDS prior use.

Final Volume (ml):           100                      50

1M Tris-HCl;pH ~8 (ml):   40                        20

0.5M EDTA (ml):              12                        6

1M NaCl (ml):                   15                        7.5

Dissolve in dd.water. Autoclave and then add 20% SDS @ 5% to the final volume.

3) Solution C    Protein ppt. buffer * Light sensitive, so store in amber bottle at 4'C; storage can be done for 1 year at the max.

This solution is 5M Sodium perchlorate= 70.23g of sodium perchlorate in 100 dd. water/ autoclaved water.

4) 0.9% NaCl (mammalian saline)

5) for 200ml TE: 2ml 1M Tris/Trizma(Sigma) + 0.4ml 0.5M EDTA. pH: 8.4

6) Cloroform (in 4'C)

7) 70% Ethanol (in 4'C)

8) Isopropanol (in 4'C)

Procedure

S1. To 1 volume of heparin added blood add 5 volumes of 0.9% NaCl. Mix by inversion. Centrifuge at 2000rpm for 10 minutes.

S2. Decant the supernatant.

S3. Add solution A- 4 vols of the original blood taken and mix slowly by inversion. The solution will turn clear red due to lysis of RBC. Spin at 3500rpm for 5 minutes. If a white pellet is clean, proceed to next step or else any red pellet indicates incomplete RBC lysis. Add 2ml of Solution A and repeat the S3. Discard the supernatant.

S4. Add 2ml of Solution B, vortex to dislodge pellet. Invert mix for 10minutes unless the pellet looses it's integrity. (To hasten the process, I suggest to keep the mix in 37'C for 5 minutes in between two inversions of 7 minutes and 3 minutes.) If the lysis of WBC takes place, the pellet looks slightly thready and the solution gains a silky appearance of khadi silk.

S5. Add 500uL of solution C and mix rigorously using vortex.

S6. Add 2ml of chilled chloroform and mix. The solution turns milky.

S7. Centrifuge at 2500rpm for 5 minutes.

S8. Transfer the clear aqous layer in 1.5mL tubes. This may take several turns to transfer all the Aq. supernatant. The suggested process:

            S8/1: Take 500uL of supernatant and add 500uL of Chilled isopropanol, Invert mix by hand. Appearance of translucent threads. These are alcohol insoluble DNA.

            S8/2: Spin down at 13000rpm for 5 minutes or 10000rpm for 12 minutes.

            S8/3; Keep the pellet (DNA) and discard the supernatant. Repeat S8/1 unless all the aqous layer from S7 spin is accumulated in one 1.5mL tube for a particular sample.

S9. Add 1ml of chilled 70% ethanol. Wait for 5 minutes and spin at the speed conditions of S8/2.

Discard the alcohol and air dry. (a maximun dry of 18 hours is allowed; overnight dry). Keep in mind, not to over dry!

Add 100uL TE to the air dried pellet. Keep in 4'C for 3 days, before estimation and analysis.

Hope this will help a lot of my molecular biologists in different arena of life science. This is a tried and tested protocol.

Tell me how to win your heart...

For I haven't got a clue-

But its always you and me and everything new!!

Five years of walking together under the welkin, its approaching a new beginning. Its calling for the violin, the cakes, celebrations and many more. But when the time for these will run down, its just me... and you!

Changing defines my life, since when I met you. From the novice walker in the park, you made me transform into a steady feet! You have pulled me out of the several ideas of changing tracks. You derailed my corporate "passion" to the real me; making me comfortable in what I did and do and will keep on doing. May be that's the clue where I won your heart and you- won it long before you even thought about winning it.

Hello.......

Its just me and you.

Story of a Little Indian State: Himachal Pradesh (Part 4)

The fourth part of the story was long due.

It was time to move out of the State Capital and romp towards the northern Himachal, 300KMs from Shimla, to a famous tourist destination called Manali. (Not Manila, as many flips their tongue!)

Manali is beautiful, no doubt, but this post is based on the beauty that the roads towards Manali that holds the lens (eye/camera), pop out of the window; throughout the journey. Residing at the foothills of the Upper Shivaliks, near the meet point of Beas, Manali is like Haridwar for Beas. For more than 70% of the journey, which took a good 6 hours in the Alto, we traced River Beas. The pebbles of the river bed executed some of the colours, close to nature.

The river had its own succulent nature. It accelerates suddenly, while slows down the flat valley. On either side lies the barren apple orchards. sibce we visit just after the apple's season, we missed the fruit from heaven, hanging, just like a common "wild" fruit around! Our driver was caring enough to tell us the geographic nature of the place and advised us that Manali and Shimla has two well defined seasons. One must visit in both the seasons, to savor the taste of Himachal. Its April-September (Apple and Sunny time) and November - February (Snow time). I hope to visit on the later part soon.

The vitality of the trip was fresh air and soothing feel. On the way, we went through a newly made tunnel, that was dark, adventorous, four lane big, bypassing underneath the Himalayas. Its 3.1Km long, but thanks to the fading memory and age catching up, I just dont remember the name.

But, the most interesting and beautiful place that I saw in this whole drive of 6 hours was 20KM away from the Manali township. Inexplicable; that's why, I allow my pictures to talk for what I mean!

Lovely!

6 years, many new things to learn!

... and I just feel I have just started; although the feeling gets interrupted abruptly, as you can see the dropping stats in my frequency in blogging out thoughts. But, guess, that's how life moves into senescence drive from adolescence! Blog too follows the rule as its life, itself!

Since 2006, my romance with this small space had started and I have tried to make it large. My history claims that I have had 1,33,200 hits since 2008, which makes a good 26.6k hits per annum. Only that my adsense doesn't pay up much as my friends only love to read my thoughts rather than search some items or products from my blog! Come-on! i am not Amazon.com! Am I? (but sometimes money also makes me miss it's presence by "selling" my thoughts)

Some simple stats:

Till now I have had posted 450+ articles. Many have had received it's comments and due share of respect, with people liking it and mostly are being channeled by Google Search and many through facebook and previously through orkut and now a days, sometimes, Twitter! I have shared some 1600+ photoes, of which 982 photoes are my own copyrighted snaps! And I flaunt them to the world. And I am not finished yet! Infact, its a long way to go: Miles to walk, before I sleep.

copyright: Somnath Paul | 2012. Bagmundi, Puruliya | WB

But, today I shall be speaking a purpose. A purpose that makes a living. A purpose that creates a living and it makes you wonder, whether you are right with or not.

Back, last week-end, I was in Bagmundi, Puruliya. Having finished my work, I managed to move out into the sun with my lens to collect some memorablia from such a serene beauty; of raw nature. A small girl came to me. In bengali, she asked: Where did I come from? Why was I there? I told her that I went there from Kolkata and I work in IICB. (She looked awed; understandably, she didnot understand what I meant in the latter part.) She asked: Was I a reporter? The reason was obviously hanging from my neck. I smile a No! I asked her why she was asking me so?

I was just dumbstruck with her reply.

In bengali she said: I thought the reporters are here and that there would be trouble between  red and green and people from both colours would come into their village to torture them, followed by the men who has camera like mine. 

To an adult Bengali, the Red and Green meant to faces of the coin at the Bengal Bidhan Sabha. That small conversation of 5 minutes left me in wonder as to what happened to these people in the past two years; thanks to the tag: Mao-baadi or Maoists! 

All are same. But, then...

Life goes on!