Thursday, December 4, 2008
advance biotech
Westerning blotting
It is used to detect the protein from the sample.
1.Protein are extracted from the sample.
2.Extracted protein are subjected to polyacralamide gel electrophorasis (PAGE).
3.Tranfer of protein from the gel to the nitrocelluse is achieved by applying a transverse electric field of 15 volt for 2 hrs.
4.Protein will bind with nitro cellulose filter.
5.Then radio labed specific antibody (prob) is added on which bind to the corresponding.
6.Autoradiograph on X- Ray is prepared.
It is used to detect the protein from the sample.
1.Protein are extracted from the sample.
2.Extracted protein are subjected to polyacralamide gel electrophorasis (PAGE).
3.Tranfer of protein from the gel to the nitrocelluse is achieved by applying a transverse electric field of 15 volt for 2 hrs.
4.Protein will bind with nitro cellulose filter.
5.Then radio labed specific antibody (prob) is added on which bind to the corresponding.
6.Autoradiograph on X- Ray is prepared.
advance biotech
DNA finger printing-
It is the identityfication of DNA sample by utilizing the particular DNA marker present of the DNA and known as mini satellite. The no. of mini sateliteis different in different sample .This difference is known as VNTR.
Satellite DNA-It is the part of DNA present on non transcribed region having repetitive sequence.
Procedure-1.A drop of blood or serum or a root of hair or virginal swap or vaginal epithelium are mainly used for the DNA finger printing.
2.DNA is extritd from the sample and is subjected to the restricted endonuclease which cuts DNA fairly frequently but not with in mini starlight.
3.These cut fragment having different different length are known as RFLP.
4.The resulting RFLPs are then electrophoresed at gel and these are blotted at nitrocellulose.
5.The minisatalight sequences present at the fragment are detected by the labled DNA probe.
6 After the hybridization of mini satelight to X-ray film.On which minisatelight region develop particular type of bands. These bands are known as DNA finger print,which are unique to each person.
The similarities and dissimilarities of these band help in the identification of person
It is the identityfication of DNA sample by utilizing the particular DNA marker present of the DNA and known as mini satellite. The no. of mini sateliteis different in different sample .This difference is known as VNTR.
Satellite DNA-It is the part of DNA present on non transcribed region having repetitive sequence.
Procedure-1.A drop of blood or serum or a root of hair or virginal swap or vaginal epithelium are mainly used for the DNA finger printing.
2.DNA is extritd from the sample and is subjected to the restricted endonuclease which cuts DNA fairly frequently but not with in mini starlight.
3.These cut fragment having different different length are known as RFLP.
4.The resulting RFLPs are then electrophoresed at gel and these are blotted at nitrocellulose.
5.The minisatalight sequences present at the fragment are detected by the labled DNA probe.
6 After the hybridization of mini satelight to X-ray film.On which minisatelight region develop particular type of bands. These bands are known as DNA finger print,which are unique to each person.
The similarities and dissimilarities of these band help in the identification of person
Friday, November 28, 2008
advance biotech
Southern Blotting-
The name of this technique is derived from the name of its inventor E.M. southern .When DNA-DNA hybridization done it is called Southern Blotting, In this technique DNA is transferred from gel to the nitrocellulose filter.
Procedure-1.DNA sample is digested with restruction enzyme.
2.Digested sample is electrophorased on agrose gel.
3.DNA bands in gel are denaturated by alkali to get single standard DNA.
4.Gel is than nutrilized by socking in tris buffer.
5.The Gel is positioned on top of buffer socked filter and a piece of transfer medium Nitrocellulose or nilone membrane) is placed on top of gel completing the gel sandwhich.
6.Dry filter paper are peeled on top of the filter and a wt put on it.
7.Capalary action pulls the buffer from the bottom filter threw the gel to the transfer medium and up threw the paper a rapid transfer by Appling vacuum also been development.
8. The arragment made for blotting ,is left undistributed for hour or over night and than top filter and wt is removed.
9.Single strand DNA fragment are traped on the transfer medium
DNA fragment fixed permanently to by baking at 80 degree centigrade nitrocellose membrane by baking with U.V. light.
10.DNA fragments on membrane can then be probed for sequence of interest by hybridization membrane is washed to remove unbound DNA.
11. X-ray film is than exposed to the membrane to get autoradiograph.
Application-
The southern blotting technique is extrely sensitive.
i.It can be used to map the restriction sites around a single copy gene sequence in any genome (even of man).
ii.It is used for DNA fingerprinting.
iii.Preparation of RFLP maps.
iv.Detection and identification of the transferred genes in transgenic individuals etc.
The name of this technique is derived from the name of its inventor E.M. southern .When DNA-DNA hybridization done it is called Southern Blotting, In this technique DNA is transferred from gel to the nitrocellulose filter.
Procedure-1.DNA sample is digested with restruction enzyme.
2.Digested sample is electrophorased on agrose gel.
3.DNA bands in gel are denaturated by alkali to get single standard DNA.
4.Gel is than nutrilized by socking in tris buffer.
5.The Gel is positioned on top of buffer socked filter and a piece of transfer medium Nitrocellulose or nilone membrane) is placed on top of gel completing the gel sandwhich.
6.Dry filter paper are peeled on top of the filter and a wt put on it.
7.Capalary action pulls the buffer from the bottom filter threw the gel to the transfer medium and up threw the paper a rapid transfer by Appling vacuum also been development.
8. The arragment made for blotting ,is left undistributed for hour or over night and than top filter and wt is removed.
9.Single strand DNA fragment are traped on the transfer medium
DNA fragment fixed permanently to by baking at 80 degree centigrade nitrocellose membrane by baking with U.V. light.
10.DNA fragments on membrane can then be probed for sequence of interest by hybridization membrane is washed to remove unbound DNA.
11. X-ray film is than exposed to the membrane to get autoradiograph.
Application-
The southern blotting technique is extrely sensitive.
i.It can be used to map the restriction sites around a single copy gene sequence in any genome (even of man).
ii.It is used for DNA fingerprinting.
iii.Preparation of RFLP maps.
iv.Detection and identification of the transferred genes in transgenic individuals etc.
Thursday, November 27, 2008
Advance Biotech
Polymerase Chain Reaction (PCR)
This technique was developed by Karymullis at Cetus corporation between 1983 & 1985.
PCR provide a simple and ingenous method for amplification of specific DNA sequence by in vitro DNA synthesis . By this technique has made it possible to synthesis the large quantities of a DNA fragment without cloning it.
The details of PCR techniques and its mechanism are described by Erlich (1989) in his edited book PCR Technology .The PCR includes the following three essential steps to amplify a specific sequence.
(i) Melting of target DNA-The target containing sequence between 100 and 5,000 base to be amplified is denatured (around 94 degree centigrade for 15 sec ) to separate its complementary strands .This process is called melting of target DNA.
(ii) Annealing of Primers-The second step is the annealing of two oligonuclotide primers to the denaturated DNA strands. Primers are added in excess and the temperature lowered to about 68 degree centigrade for 60 sec. Consequently the primers from hydrogen bonds i.e. anneal to the DNA on both sieds of the DNA sequence.
(iii) Primer Extention-Finally, nucleoside triphosphate (dATP,dGTP,dctp,dTTP) and a thermo stable DNA polymerase are added to the reaction mixture.The DNA polymerase are added to the reaction mixture.The DNA polymerase accelerated the polymerization process of primers and therefore,extends the primers (at 68 degree centigrade) resulting. In synthesis of copies of target DNA seq. only those DNA polymerase which are therstable i.e. ucton at the high temperature are employed inPCR technique.
For this two popular enzymes are used Taq polymerase ( a thermophilic bacterium thermos aquticus) and vent polymerase-Thermococcus litoralis are used in PCR technology.
Theses enzymes exhibit relative stability at DNA melting replacement after each cycle of synthesis.
Also it reduces the cost of PCR and allow automated thermal cycling.
After completion of step 3 (of one cycle) the targated sequences on both strands are copied and four and four strands are produced. Now the three step cycle is repeated which yields 8 copies from four strands.
Similarly, the third cycle produces 16 strands. This cycle is repeated about 50 times.
20 cycles (each of three steps) will produce about one million copies of the target sequence 30 cycle will produces one billion copies.
For working of PCR about 10-100 picomoles of moles are required.
The con of target DNA can be about 10-20 to 10-20 M or 1-105 DNA copies per Ml.)
PCR machine can carry out 25 cycle and amplify DNA 10 5 times in 75 min. The rTth DNA polymerase will transcribe RNA to DNA. Thereafter amplify the DNA.
Therefore, cellular RNA and RNA virsues may be studied when they are present in small quantities.
Application of the PCR Technology-
1.Amplification of DNA and RNA.
2.Determination of orientation and location of restruction fragments relative to one another.
3.Diagnosis of disease and causal micro-organisms.
For example- PCR based diagnosis test for AIDS,Chlamydia, tuberculosis,hepatitis.
4.The PCR is important in detection of genetic disease such as sickle cell anaemia Phenyketonuria and muscular dystrophy.
5.It is also applied in diagnosis of plant disease. Alarge no. of plant pathogens in various hosts or environmental sample are detected by using PCR, for example viriod.
This technique was developed by Karymullis at Cetus corporation between 1983 & 1985.
PCR provide a simple and ingenous method for amplification of specific DNA sequence by in vitro DNA synthesis . By this technique has made it possible to synthesis the large quantities of a DNA fragment without cloning it.
The details of PCR techniques and its mechanism are described by Erlich (1989) in his edited book PCR Technology .The PCR includes the following three essential steps to amplify a specific sequence.
(i) Melting of target DNA-The target containing sequence between 100 and 5,000 base to be amplified is denatured (around 94 degree centigrade for 15 sec ) to separate its complementary strands .This process is called melting of target DNA.
(ii) Annealing of Primers-The second step is the annealing of two oligonuclotide primers to the denaturated DNA strands. Primers are added in excess and the temperature lowered to about 68 degree centigrade for 60 sec. Consequently the primers from hydrogen bonds i.e. anneal to the DNA on both sieds of the DNA sequence.
(iii) Primer Extention-Finally, nucleoside triphosphate (dATP,dGTP,dctp,dTTP) and a thermo stable DNA polymerase are added to the reaction mixture.The DNA polymerase are added to the reaction mixture.The DNA polymerase accelerated the polymerization process of primers and therefore,extends the primers (at 68 degree centigrade) resulting. In synthesis of copies of target DNA seq. only those DNA polymerase which are therstable i.e. ucton at the high temperature are employed inPCR technique.
For this two popular enzymes are used Taq polymerase ( a thermophilic bacterium thermos aquticus) and vent polymerase-Thermococcus litoralis are used in PCR technology.
Theses enzymes exhibit relative stability at DNA melting replacement after each cycle of synthesis.
Also it reduces the cost of PCR and allow automated thermal cycling.
After completion of step 3 (of one cycle) the targated sequences on both strands are copied and four and four strands are produced. Now the three step cycle is repeated which yields 8 copies from four strands.
Similarly, the third cycle produces 16 strands. This cycle is repeated about 50 times.
20 cycles (each of three steps) will produce about one million copies of the target sequence 30 cycle will produces one billion copies.
For working of PCR about 10-100 picomoles of moles are required.
The con of target DNA can be about 10-20 to 10-20 M or 1-105 DNA copies per Ml.)
PCR machine can carry out 25 cycle and amplify DNA 10 5 times in 75 min. The rTth DNA polymerase will transcribe RNA to DNA. Thereafter amplify the DNA.
Therefore, cellular RNA and RNA virsues may be studied when they are present in small quantities.
Application of the PCR Technology-
1.Amplification of DNA and RNA.
2.Determination of orientation and location of restruction fragments relative to one another.
3.Diagnosis of disease and causal micro-organisms.
For example- PCR based diagnosis test for AIDS,Chlamydia, tuberculosis,hepatitis.
4.The PCR is important in detection of genetic disease such as sickle cell anaemia Phenyketonuria and muscular dystrophy.
5.It is also applied in diagnosis of plant disease. Alarge no. of plant pathogens in various hosts or environmental sample are detected by using PCR, for example viriod.
Sunday, August 3, 2008
Anthurium-Introduction-2
The Anthurium is an ‘evergreen’ which can produce flower all year round .Northeast India, with its diverse climatic conditions from tropical to temperate/alpine zone is an ideal region for floricultural activity. Anthurium International Including currently offers nearly 800 variations of tropical flowers, foliages and plants. However, in spite of all congenial agro-climate and abundance of genetic resource the region has remained backward in the floricultural sector- especially Anthurium orchids. This is mainly due to the infrastructural constraints in communication (of all types), difficult terrain, inaccessibility, inadequate R&D support etc.
In 2005, the Anthurium took ninth position and the Anthurium pot plant was ranked forth in the cut flower in Dutch auctions listing with 11.1 million supplied , an increase of 1% over the previous year and average price was uro 3.58.
Demand of Anthurium flowers are 38%. There are excellent scope to Japan west Asia, Singapore and EU countries. Now the demanding rate is grow about 25-30 % per annum.
Right from the British days, Uttaranchal has been involved in floricultural activities because of the ideal agro climate with varying altitude, temperature and rainfall regimes. Anthurium has been cultivated by some hobbies and nursery men over the last half century in a traditional and natural manner for the trade of plants only. However in the recent years, recognizing the potentials of Anthurium in export oriented economic activity, the state government is trying to promote Anthurium based floriculture in the state. The centre has contributed significantly in developing the cultivation packages and promoting floriculture as a supplemental crop in the villages. However, the state is yet to develop a model Hi-Tech Anthurium farm for export oriented production & trade meeting the international standards.Hence, the present project proposal envisages the establishment a Hi-Tech model Anthurium farm which would serve as a nucleus of activities in promoting hi quality production in a large scale involving the local farmers women & youth and developing subsequent marketing support.
In the recent years however, under the Priminister’s package for development of NE region, some infrastructure is coming up. Uttaranchal, Guwahati & Kolkata airports are busy modernized to handle floricultural product aiming at export development with cold storage facilities .futher, Floriculture is reciving some attention encouraging the entrenevers in a small way. North Eastern council , Shillong is also playing a vital role in the over all development of this region.
Following are specific objectives of the project
Ø To encourage the farmers of Rudrapuare for Anthurium cultivation under GreenHouse conditions.
Ø To fulfill the demand of Anthurium in International and Domestic Market.
Ø Provide the employment for unemployed youths,Women and farmers in Kumaon region..
Ø Provide all the facilities for the production of Anthurium in Rudrapur, as a production centre.
Ø To train the villagers for Anthurium cultivation.
Ø Establishment of cold storage unit and promoting export at national & International level.
In 2005, the Anthurium took ninth position and the Anthurium pot plant was ranked forth in the cut flower in Dutch auctions listing with 11.1 million supplied , an increase of 1% over the previous year and average price was uro 3.58.
Demand of Anthurium flowers are 38%. There are excellent scope to Japan west Asia, Singapore and EU countries. Now the demanding rate is grow about 25-30 % per annum.
Right from the British days, Uttaranchal has been involved in floricultural activities because of the ideal agro climate with varying altitude, temperature and rainfall regimes. Anthurium has been cultivated by some hobbies and nursery men over the last half century in a traditional and natural manner for the trade of plants only. However in the recent years, recognizing the potentials of Anthurium in export oriented economic activity, the state government is trying to promote Anthurium based floriculture in the state. The centre has contributed significantly in developing the cultivation packages and promoting floriculture as a supplemental crop in the villages. However, the state is yet to develop a model Hi-Tech Anthurium farm for export oriented production & trade meeting the international standards.Hence, the present project proposal envisages the establishment a Hi-Tech model Anthurium farm which would serve as a nucleus of activities in promoting hi quality production in a large scale involving the local farmers women & youth and developing subsequent marketing support.
In the recent years however, under the Priminister’s package for development of NE region, some infrastructure is coming up. Uttaranchal, Guwahati & Kolkata airports are busy modernized to handle floricultural product aiming at export development with cold storage facilities .futher, Floriculture is reciving some attention encouraging the entrenevers in a small way. North Eastern council , Shillong is also playing a vital role in the over all development of this region.
Following are specific objectives of the project
Ø To encourage the farmers of Rudrapuare for Anthurium cultivation under GreenHouse conditions.
Ø To fulfill the demand of Anthurium in International and Domestic Market.
Ø Provide the employment for unemployed youths,Women and farmers in Kumaon region..
Ø Provide all the facilities for the production of Anthurium in Rudrapur, as a production centre.
Ø To train the villagers for Anthurium cultivation.
Ø Establishment of cold storage unit and promoting export at national & International level.
Saturday, August 2, 2008
Anthurium-Introduction-1
Anthurium are one of the most popular tropical with long vase life of about six weeks and even more depending in the variety and season Anthurium are herbaceous epiphytes, native to tropical America. The Anthurium is also known as painted Tougue, Flamingo flower ( Flamigo lily) or Tail Flower. Anthurium are grown for their brightly colored flower spathes and their ornamental leaves.Family Araceae which includes more than 100 genera and about 1500 species. The Anthurium is perennial herbaceous plant usually cultivated for its attractive , long lasying flowers . This flower is preferred for their colourful, modified leaf (spathe) and hundereds of small,Botanical flowers on the pencil like protrusion (spadix) rising from the base of the spath. The Anthurium plant produces flowers throughout the year. One flower emerages from each leaf axil. The sequence of leaf, flower and new leaf is maintained throughout the entire life of the plant. The interval between leaf emergences is shortened or lengthend with changes in environmental conditions during summer more flowers can be expected per plant than during the winter when there is less light and lower temperatures.
Anthurium orchids is one of the important floricultural crops for the Northeast India, Uttaranchal in particular Kumaon regoin. Amongst the Orchids, Anthurium command a place of pride and one of the important foreign exchange earners because of their long lasting variously colored attractive flowers arranged in erect stems.
Anthurium are member of the Arun lily family (Araceace) and sub family Pothoideae & the order of the Anthurieae . Greek words ‘ Anthos’ and ‘Oura’ which means , ‘Bloom’ and ‘Tail’ respectively.
Anthurium often refered to as Flamigo Flower or tail flower, Anthorium are an exotic bloom with long , waxy petals with in the centre. Though by many as the ‘Hawaiian Flower’ This species is found in their natural habitat only Mexico, Central America and the West Indies. Anthurium were introduced in to Asian rain forest and Hawaii and can be founded growing wild there todays.
Anthurium orchids is one of the important floricultural crops for the Northeast India, Uttaranchal in particular Kumaon regoin. Amongst the Orchids, Anthurium command a place of pride and one of the important foreign exchange earners because of their long lasting variously colored attractive flowers arranged in erect stems.
Anthurium are member of the Arun lily family (Araceace) and sub family Pothoideae & the order of the Anthurieae . Greek words ‘ Anthos’ and ‘Oura’ which means , ‘Bloom’ and ‘Tail’ respectively.
Anthurium often refered to as Flamigo Flower or tail flower, Anthorium are an exotic bloom with long , waxy petals with in the centre. Though by many as the ‘Hawaiian Flower’ This species is found in their natural habitat only Mexico, Central America and the West Indies. Anthurium were introduced in to Asian rain forest and Hawaii and can be founded growing wild there todays.