This study regards the 18 new varieties ofs which introduced as the examination material, on three different soil condition such as upland and brown soil, terraced fields and mucosity soil in the mountain fields, bottomland and sandy soil, we comprehensive appraised the 18 new varieties of s about the phenophase, growth characteristics, early period fruit characteristic, quality of nut and resist characteristic on adverse conditions etc.. We studied the Chinese chestnuts red spider hurt the different varities and the situation of output decrease using cluster analysis. Also we described and summarized the character botany, the biology speciality about some choiceness varieties which has development future. We have pertinent bring our development suggestions, the main result is as follows: 1.Through studied on the phenophase, growth characteristic, fruit characteristics, chestnut real quality and resist characteristics on adverse conditions of the 18 Chinese chestnut varieties which introduced, we screened out Hou84-3, Dal-3, Yanhong which are the varieties that suitable to extended in Tai’an and middle of Shandong Province area.These three varieties which in 3 years old have more than 1.8kg produced of average individual plant, have more than 2.5kg produced of average individual plant,they are fertility and constant-rate of production. Single nut weight is more than 8.5g. The integration characteristics are excelled than Shifeng which had already applied in yield.In addition Yanshanduanzhi has coppice characteristics obviously, this variety could conveniented for intensivism producded and its nut have best qualities, this variety can resist drought conditions. can developed properly in the fertile mountain region terraced fields of soil.2.On the basis of consulting documents, according to the Chinese chestnut breeding experts and the growth expert’s suggestion, we regard the merchandise characteristics as the most important factor, we has gone on onthe comprehensive appraisal of the real quality of every chestnut variety, the result is:The 5 varieties Yanshanduanzhi, Dal-3, Hou84-3, GuantinglOhao, Yanhong have best chestnut quality. The Yankui, Shifeng, yanchang, 10-14 have moderate quality of Chestnut. The other varieties such as Qianxi2399 has bad characteristics.3. In the drought soil conditions, we mensurate the photosynthetic rate and other mutuality index, the growth characteristics and per mu yield. We analysised the drought resisted characteristics of all the Chinese chestnut, the result is, Yanhong, Yanshanduanzhi and Hou84-3 can bear stronger drought conditions .Yanhong cann’t bear drought conditions.4. We dicussed the hurt and the yield reduce about Chinese chestnut red spider harm on different varieties. We regard the degree of lamina losing green as the index, using the method of cluster analysis,the result is that the 9 varieties such as Yanshanduanzhi, Hou84-3 etc. have light hurt on Chinese chestnut red spider.The Yankui , Chushuhong is endangered by the chestnut red spider seriously.5. We described the character botany, the biology speciality such as lamina, chestnut shed, thorn, chestnut and autumn etc. of some choiceness varieties which has development future.Find that the lamina, chestnut shed, thorn, chestnut and autumn etc. are the critical characteristics which can distinguished these varieties.
Aim at the current situation of serious environmental pollution caused by intensive husbandry, the author discussed the necessity and possibility of realizing organic agriculture in China, and elucidated that disposal of livestock manure toland is the most economical and environmental sound solution for organic agriculture. was required to estimate the nutrient contents in animal manure for utilizing animal manure as a resource. There are two methods for estimating the nutrient contents in animal manure. The first one is rapid tests based on the testing technology of physical- characteristics, and the second one is mathematical model. This paper reviewed the two methods separately and discussed the further study at the same time. Quick tests could estimate the nutrient contents rapidly by constructing the relationships between the physical- characteristics and total nitrogen, phosphorus~ potassium and ammonium contents in animal manure, and then testing these physical-mechanical characteristics, such as specific gravity, conductivity and dry matter, etc. According to some independences, such as dietary intake, liveweight, etc, the nutrient contents in animal manure could be estimated rapidly and accurately through the calculation of the mathematical model. The two methods, which are more convenient, rapid and low-priced, compared with the conventional lab analysis, will save a great lot of human and material resources when put them into practice, and have important theoretical and applied values for promoting to use animal manure scientifically and fulfilling the sustainable agriculture. Studied the mathematical model method, which is based on the data of poultry and the formulation of their dietary intake. By theoretical analyzing the influential factors of the volume and nutrient contents of animal manure, the independences of the model were selected and they were animal type, age, liveweight, feed input, dry matter intake, N, P and K intake, dry matter output and slurry output. Broiler -Iv – experiment was carried out from which a large number of data were obtained. By regression analysis, the most important factors for estimating the volume and N, P, K contents of broiler manure were attained. A series of empirical model were developed. was tested that these models were reasonably reliable. Using animal age as a predictive independence was presented for the first time. Simple equations of using broiler age as an independence were developed separately, and the regression relationships were highly significant. Combining age and other factors instead of liveweight attained multiple equations, which are more reliable and useful. Selected the most reliable, economical, simple and useful models. The results of the estimation were identical with the experiment results on the whole. Studied the rapid tests method, which is based on the testing technology of physical-mechanical characteristics. 20 broiler slurry samples were collected for the purpose of studying the relationships between specific gravity, conductivity and the nutrients in animal manure. According to the results of regression analysis~ specific gravity can estimate the P contents in animal manure, and conductivity can estimate the ~ and K contents precisely. Besides, the influence of dilution to the conductivity of the solutions was tested. The
Bivalves constitute a dominant and divers group of marine animals in our country. Most of them are of major commercial important species, and studies on theirare necessary for the sustainable exploitation and conservation of these bioresources. The objective of the present study is to explore the feasibility of using the as a molecular marker for studying the interspecific phylogenetic relationship and intraspecific genetic variations among . The 18S-ITS1 sequences of 11 species at differing taxonomic levels were determined. The sequences were found to exhibit a high degree of length polymorphism among different species, ranging from 558 bp (Chlamys farreri) to 784 bp in (Ostrea rivulahs), mainly resulting from the variation of ITS1. The percent divergence of sequences ranged from 3.8% to 60.4%. The NJ (neighbor-joining) tree inferred from 18S fragment of 14 species agree with the previous based on morphologies and chemical analysis well. The sequence was found to vary among 4 geographical sites of Pinctada martensi, ranging from 0.6% to 1.9%. The variations involved substitutions, transversion as well as insertions and deletions among them in P.martensi. All these results show that ITS1 is highly divergent among different species of and could be used in classification and distinguishing closely related species, and also could be used for molecular systematic studies at relative species, subspecies and population levels.Pinctata martensi is one of the most important aquacultural sea s in the southeast of China. Wild individuals were collected from Sanya (Hainan province), Beihai (Guangxi province) and Daya Bay (Guangdong province) randomly, then mated and cultured in the same condition. In order to get a deep understanding of and improve efficiency of breeding by marker-assisted selection, we using 27 primers tested different filial populations by technique. The results showed that the diversity Shannon index of filial population mated from inter-populations was higher than filial populations mated from inner-populations. Then we analyzed the feasibility of breeding from two different areas.
, one kind of western thick-skin melon, has high economic value, because of its characteristics of very special flavor, fine texture, and good taste. For the demand in Ningbo, we introduced new high-quality varieties of muskmelon from Japan. Through two years field trials, some good varieties such as “Earl’s darling” (ァ一ルスダ一リソ) have been successfully selected. In order to improve the cultivation techniques, the experiments have been carried out to systematically study on variety suitability, fertilization, disease and pest control, , etc. and their influences on quality.The results of variety suitability experiments in the seasons of spring, summer and autumn suggested that the variety “Earl’s darling” was obvious superior to other varieties and was suitable for protected cultivation in Ningbo and surroundings. “Earl’s darling” had characteristics of developed root system, strong main stem and leaves, relatively large and round shaped fruits covered with light grayish green and heavily netted surface. needed about 59 and 56 days from blooming to fruit ripening in spring and autumn respectively. The average weight per muskmelon was 1.8 Kg wi th about 3. 9 cm thick pulp. The fruits had higher hardness, the flesh was yellowish green, flavorful, delicious, juicy and crisp, and the central sugar content was about 15. 5%. This variety was well adapted, tolerant to low temperature, resistant to the powdery mildew and relatively low incidence of gummy stem blight. The whole growth period was about 135 days in spring. was easy to fruit set, planting manage, storage and transport. The experiments showed that all fruit quality indexes of “Earl’s darling” were better than control variety “Shengtian No. 1″, among which vitamin c was 2.78 times of control, the contents of mineral nutrients such as iron, calcium etc. were much higher than the control variety and reached by 38.6%, 13.9% respectively, and the content of ash and protein were 3.39% and 24.1% higher than the control.Cultivation techniques experiments showed that the available operation for “Earl’s darling” were single-vine , one fruit in one plant, top removing when the 25th true leaf emerged in main vine, better fruit setting in the 13th-15th nodes in spring and in the 12th -14th nodes in autumn. Field survey further indicated that the single-vine with one fruit per plant or double-vine pruning with two fruits per plant was suitable in spring, and the single-vine pruning with one fruit per plant was suitable in autumn, but in summer, the double-vine pruning with one fruit per plant was more suitable showing that the net was beautiful, fruit size was medium, the sugar degree was high, and the yield was near to the output of which the single-vine pruning with one fruit per plant. The time of sowing was from July 10th to August 10th for autumn cultivation in Ningbo, and it couldn’t be later than the August 15th. Using the method of (the medium was made of sawdust with a little of thoroughly fermented manure and added 25% of the soil, and the proportion of firm phase, liquid phase and gaseous phase were 20%, 40% and 40% respectively), the plant had grown vigorously and couldn’t tend to per-senescence, the net was pretty, the fruit was easy to enlarge and had high sugar content and good quality, the yield was remarkably improved, the economic profit was outstanding, and at last the integrated disease resistant was notable enhanced. The “Yong stock No.3″ and “Earl’s darling” showed the best compatibility, and the grafted seedlings were better than the self rooted seedlings. Through combined with single-vine pruning, plants were effectively prevented from soil-born diseases such as fusarium wilt etc., and withered phenomenons caused by toxic acids or low oxygen around roots. In order to effectively control the incidence of gummy stem blight, some measures could be taken such as reducing air relative humidity, pruning, removing old leaves in time, and more important, daubing “(?)traconazole (?)carbendazin” on the stem combined with spurting “tetraconazole” in the whole plant. In addition, 20% 2000×”Yizailing” (a new type of plant growth regulator) was watered to strengthen seedling and early recover during the period of seedling and transplanting, and also to improve plant resistant to fusarium wilt.
Through systematic studies on techniques used in sheep and?embryo transfer and their affecting factors, we have reached the following results: 1. There is no significant difference in available embryos between natural estrous donors and P0F21 induced estrous donors(p>0.05), but available embryos decrease significantly after super-ovulation operation by using CIDR to donors(p<0.05). 2. Two FSH preparations are used in sheep. and the average embryos collected from each group of donors are: 12.85? 6.99(NingBo, 1501U), 12.4?5.74(NingBo,1801U) 9.32 ?4.83(CAS. 2.5mg), 11.71 ?4.53(CAS, 3.0mg). Carefully using of the two preparations can have good results in super-ovulation. 3. The best season for sheep super-ovulation in Inner Mongolia is between the middle of September and the middle of November, the best season for Boer goat super-ovulation is between the middle of October and the Late of November. 4. Same dosages of FSH have no significant effects on Germany mutton merinos and (P>0.05). 5. The age of donor sheep has no significant affection on super-ovulation. First time induced ovulation to donor sheep soon after sex maturity with adequate lower dosage can reach same result as good as ewes which already have lambs. 6. There is no significant difference in average ovulation rate between replicates of super-ovulation to donor sheep (p>O.O5), but there is a significant difference in available embryos collected(p,0.01). 7. Average available embryos from sheep and under supplement are significantly higher than those without supplement (p<O.O 1). 8. The best time to collect embryos for producing frozen embryos from donors is 6.5 days during estrous. (. Good nutrition and managementis the key of pregnancy rate after embryos are transferred, lambingrate of recipients under supplement (8l %) is significantly higherthan that without supplement(76. 1 %). 0. Time difference of estrousbetween donors and recipients is very important to the lambing rateof recipients, pregnancy rate at time difference of 0h, f l2h issignificantly higher than that of 1 24h(p<0.01 ). 11. Pregnancy rateand lambing rate of the recipients transferred with 2-3 embryos aresignificantly higher that transferred with single embryo (p<0.01 ).Pregnancy rates are as fol1ows: 65. 12%(single), 79.4(double),86.2%(triple). 12. Pregnancy rate of ewes transferred with frozenembryos(66%) is significantly higher than that of ewehogg(5 5 %)(P<0. 0 1 ).
Interest subsidy loan project (ISLP) which has played a positive role in expanding industrial scale, adjusting industrial structure and prolonging the industrial chain in forestry has been implemented for 23 year in Shaanxi Provinces. Theon the practice and experience of ISLP in forestry and the exploration in how ISLP supports the forestry industry will be obviously important to the forestry work at present and also of great reference to the forestry policy-making.Focus on the current situation that the loan scale is too small and is difficult to loan, illustrate the background of ISLP in forestry and propose the objective and significance of this . Based on the description of the sustainable development theory, institutional change theory, community forest theory, investing and financing theory, propose that raise the micro-credit to support ordinary farmers to promote their capacity building. The key point of improving the environment is the effective institutional arrangement and policy measures. According the policy and theory of ISLP of our country, the function and effection of ISLP in ChannXi, analyse the problems and infault and reform.The ISLP serves the reform of forest property right which can imrove the ISLP in a more large scale. Centered on the forest- right reform and forest industry development, illustrate that changing the concept of ISLP, multi-aim developing , paying attention on the benefits and desire of farmers and innovation meathod ,new products of credict loan. By the way of which promote the farmers activity, solve their fund ineffeciency in order to develop forestry industry, construct a management system which centered on the supportting the key company of frest based on the company leading forest base and forest base leading farmers considering all influce factors. Innovating a new system which support farmer and forest staff by ISLP which can advance the forest-right reform, promote the ISLP by forest-right mortage. Suggest improving and perfecting surport policy to key company by identifing year-interests and inducting interests according to forest cycle, manufacture types; for forest micro-loan should accord to three patterns, one is basing on the key company—-”company + forest base + foerst farmer”, the another is double ways loan—-”bank+intermidiary group+forest farmer”, and the third is loan by rural credict bank. Illustrat that the key of forest property management is evaluating mortage property, keeping watch the risk, supervising the management and paying the loan. Propse five patterns of loan by foret property certificate mortage in order to solve the forest farmer mortage loan problems.The paper has proposed strategies and suggestions to innovate the policy of in forestry, pointed out the methods to improve , the necessity and importance to making good use of forest property right mortgage loan and expand the microcredit in forestry.
Jining Grey goats that has significant characteristics of highand nonseasonal ovulatory activity is an excellent local goats breed in P.R. China. The kidding percentage averaged 293.65 percent in Jining Grey goatss. Prolactin receptor (PRLR) gene which controls the fecundity of Belclare and Cambridge ewes was studied as a candidate gene on the of Jining Grey goats. According to the sequence of ovine PRLR gene, two primer pairs were designed to detect single nucleotide polymorphism of intron I of PRLR gene in both high fecundity goats breed (Jining Grey goats) and low fecundity goats breeds (Boer goats, Wendeng dairy goats, Liaoning Cashmere goats, Beijing native goats) by - . The products amplified by primer 1 had polymorphism. Five genotypes (AA, AH, AK, HH and HK) were detected in Jining Grey goats, and two genotypes (AA and AK)in other four goats breeds.The sequencing results indicated that there was one single nucleotide mutation (C~119→T~119) at intron i of PRLR gene in Jining Grey goats.. Frequencies of A, H and K alleles were 0.853,0.0759, and 0.0714 in Jining Grey goats, respectly. The PRLR genotype distributions were high significantly different (P<0.01) between Jining Grey goats and Boer, Wendeng dairy goats, Liaoning Cashmere goats. The ewes with genotype AA had 0.6 (P<0.05) kids more than those with others genotypes in Jining Grey goats. The products amplified by primer 2 had polymorphism. Three genotypes (CC, CD, and DD) were detected in Boer goats, and two genotypes (CC and CD)in other four goats breeds.The sequencing results showed that there were two nucleotide mutations (G~35→A~35, and A~86→G~86) at intron II of PRLR gene in goats. Frequencies of C and D alleles were 0.9375 and 0.0625 in Jining Grey goats, respectly. The PRLR genotype distributions were high significantly different (P<0.01) between Jining Grey goats and Boer goats, Wendeng dairy goats, Liaoning Cashmere goats. The femails with genotype CC had 0.66 (P<0.05) kids more than those with genotype CD Jining Grey goats.These results implied that PRLR gene is associated with a major gene influencing the of Jining Grey goats. In view of our results, marker-assisted selection using PRLR gene is warranted to increase litter size in goats and will be of considerable economic value to grey kidskin and mutton producers.
Research on Technology of Comprehensive Prevention and Cure and Infectious Serositis to the Influence in Produce and Economic Performance of Meat Ducks
This research passes the meat duck Infectious Serositis that the proper and good county takes place in the last few years to produce and theof the economic performance with technical study of prevention and cure, affects ducks of seven-day-old-over age,especially of fifteen～twenty-day-old age,mortality was ten～seventy percent.In the acute form, listlessness,neck tucked in,ocular and nasal discharges,diarrhea,incoordination, tremors of the head and neck,are commonly seen.The chronic form was a twisted neck.Typical lesions found in post-mortem bird were infected fibrinous pericarditis, perihepatitis,airsacculitis,and septicaemia.The pathogenic germ was easily isolated from liver,spleen and blood,especially from brain tissue.The detail physiological and biochemical identification,serotype identification and drug senitive test were done on thirty-eight strains. showed that all strains didn’t ferment glucose,maltose, fructose,lactose,mannitol,didn’t disintegrate urea,didn’t produce hydrogen sulfide, didn’t deoxidize nitrate,catalase appeared positive,and could liquefy gelatine.The drug senitive test indicated that Riemerella anatipestifer(RA)was sensitive to ampicillin,cefalotin sodium,cefalexin,levofloxacin,apramycin sulfate,was insensitive to streptomycin,leucomycin.Its serotype was identified to be I by agglutination reaction.Vaccine,Chinese herbal medicine and Western medicine were applied to prevent and cure .There hundred and fifty ducklings were allocated randomly to six treatment groups and one control group with two replications:Each group was dealed with varied prescription:YajiangningⅠ,YajiangningⅡ, YajiangningⅠ+vaccine,YajiangningⅡ+vaccine,vaccine,and Western medicine+ vaccine were applied to treatments T1,T2,T3,T4,T5,andT6.Control group T7 was without any prescription.The result demonstrated that mortality of T1,T2,T3,T4,T5, and T6 was 8,6,8,4,10,13,and 22%respectively,T1,T2,T3,T4,were significantly lower than T6(P＜0.01);T1,T2,T3,T5 were not different compared with T4; difference between T4 and T6 appeared significant;T4 had the best effect on preventing and curing .At the same time,the cured rate could reach 70%by clinical cure;Different experiment the set to the meat duck produce and the economic performance all has different degree of ,but the difference does not show(P＞0.05)In general,the application to Chinese herbal medicine+ vaccine,coordinating enough preparation before breeding and careful management practices were highly effective for prevention of infection,and was valuable for use in ducks.
The Study on Soil Microbial Ecology in Taxus Chinensis (Pilg.) Rehd Plantation of Chinese Nature Reserve
Taxus belonging to the national I orⅡlevel of protection plant, and is also a rareand endangered plant in China. There are 14 kinds in the world, which our countryhas 4 and one variety, T.chinesis（Pilger）Rehd is a Chinese endemic species, which isthe national one level of protection plant. They were born in the altitude of10001200m mountainous regions. Taxus is the plant of a fine material used formedicinal use and the botanical garden tree seed. Thushas a high value formedicinal, sightseeing, scientific research and developing use. XiaolongshanNational Nature Reserve is located in west the Qinling Sierra, and upper branch ofthe Jialing River upstream, is a subtropic transitional area forest ecosystem. is inthe western area, also one of the important ecology shelter for zoology and botanyresources. This region is suitable for the Taxus chinensis （Pilg.） Rehd. Therefore it isnecessary to understand the Taxus chinensis （Pilg.） systematically andcomprehensively and the forest itself the relation between ecology and theenvironment factors, in order to use and protect this precious resource to providefundamental theory and the technical guidance for further development. Now themajor results studied in the paper are as follows:1. The soil was slightly moist soil; pH is from 6.3 to 7.2. It was basically thelow-acid. This is the typical forest soil; the soil fertility is highs. Soil organics is5.64%8.66%. This region is a high-fertility area; the entire N content is quite highand concentrated, the fast-effect N content difference is great; the entire P content isquite concentrated. The fast-effect P content belongs to the medium level basically;the entire K content is also high. The fast-effect K content ranging scope is large.2. The total quantity of soil microbe of surface layer（020cm） is 10.3×106ind./g·dry soil, which under layer（2040cm） is 4.07×106ind./g·dry soil That’s thesurface layer is lower level 2.53 times. Bacterium occupies the overwhelmingsuperiority. Each kind of microorganism’s quantity increases with the soil layer depthreducing. The quantity of soil microbe in different sampling points varies greatly. 3. The community of soil fungi is mostly imperfect fungi and zygomycete. Thedominant fungal genera are Penicillium and mucor. The familiar fungi genera areAbsidia，Trichoderma and Aspergillus.The community of soil actinomyces is mostly imperfect Streptomyce. Theyare Albosporus, Flavus, Roseosporus, Glaucus, Cinereus, Griseorubroviolaceus,Griseofuscus, Aureus, Viridis, Hygroscopicus, Micromonospora and Microbisporaare the rare actinomyce genera.4. The quantity of the soil is main physiological group increases with the soillayer depth reducing. The overall performance is: aminate > bacteriumazotobacter >cellulose decomposing bacteria.5. SOC, MBC, MBN and the respiratory rate reduces with the soil layerdeepening. The MBC/SOC value rises along with the soil layer, which showes thatthe soil carbon is in the process of during the accumulation. Soil microbialrespiratory rate is generally from 0.59 to 0.96 mg·kg-1·h-1in 020cm, and from 0.21 to0.60 mg·kg-1·h-1 in 2040cm, qCO2 is from 0.76 to 1.21 mg·g-1·h-1in 020cm, andfrom 0.75 to 1.11 mg·g-1·h-1 in 2040cm.6. The species richness index of soil fungi is from 2.4 to 3.37. Evenness index isfrom 0.38 to 0.66. Sampling field 3 has the highest diversity index H’and D whichsampling field 7 the lowest. The species richness index of soil actinomyce is from1.21 to 2.41. Evenness index is from 0.49 to 0.69. Sampling field 4 has the highestdiversity index H’and D, and sampling field 3 and 5 the lowest.7. Correlation analysis indicates that there exists a significant or highlysignificant relationship between the soil microorganisms and soil pH, soil moisturecontent, soil organic content. Soil microbial biomass has a significant relationshipwith soil organic matter contents. Soil respiration and q（CO2） have a significantrelationship with available nutrient. There is a certain relationship between soilmicrobial biomass and soil respiration. MBC has a highly significant positiverelationship with SOC. MBN has a highly significant relationship with soil respiration and q（CO2）. There is a complex relationship between soil microbialdiversity and ecological factor.
fleck disease caused by grapevine fleck (GFkV) is a worldly distributed disease. In China, the occurence of grapevine fleck disease was reported in Liaoning, Shangdong and Xinjiang. Usually the disease doesn’t show any visible symptom on most grapevine cultivars and their rootstocks. Establishing a rapid method to detect GFkV is very an important role for the controllingof this disease.In this study, serological methods were employed to detected GFkV in 10 grapevine samples collected from the Fruit Tree Research Institute of Chinese Academy of Science. The results showed that:there were four, two, one and two samples infected respectively by Virus B (GVB), FanLeaf Virus(GFLV), Grapevine Leaf Roll-associated Virus-1(GLRaV-1) and Grapevine (GFkV). Protein A Sandwich Enzyme Link Immunosorbant Assay (PAS-ELISA), Plate Trapped Antigent-Indirec-ELISA (PTA-I-ELISA) and Triple Antibody Sandwich (TAS-ELISA) were respectively employed for the GFkV assay in grapevine phelome extraction solution in different dillution, the postive result was obtained when the crude of GFkV of the infected sample in the bark was diluted respectively by 1:200; 1:500; 1:2000. Both indirect Tissue Blot Immunoassay (DTBIA) and Dot-Immunobinding Assay (DIBA) were successfully applied to detect GFkV in the petiole and leaves, both have the advantages that the results were visual and less time was neededIn addition, Immunosorbant Electromicroscope (ISEM) was applied to GFkV detection in Shengbao sample, some spherical particles were founded, these particles were decorated by the polyclonal antibody of GFkV. These results indicated that the spherical particles were GFkVAn Immunocapture-ReverseTranscript-Polymerase Chain Reaction (IC-RT-PCR) was employed to detect GFkV in Shengbao, An about the 380bp fragement was amplified and was longer than expected size