鸟类剥制标本缺少羽毛部位的修补

在制作鸟类剥制标本时,常常会遇到鸟体某个部位因缺少羽毛覆盖而露出皮肤,给人以美中不足之感。为弥补这个不足,可以从相同鸟类的另一只鸟体上摘下同部位的羽毛(羽毛颜色,羽片大小等应尽可能相似),将羽根部剪去适当长度以后,用市售乳白胶粘贴到剥制标     

鸟类假剥制标本的制作

对于保存于制的鸟类皮肤和羽毛,主要通过制作假剥制标本来实现。假利制标本又称冰棒标本（供从事鸟类科学研究工作实用的、可作为科学资料保存的标本）,是鸟类研究的参照物,也是大多数自然历史博物馆和大学生物系用于展示和教学而收集的对象。鸟类的体型多种多样,但制作方法基本一致。工工具及材料解剖盘、解剖刀、剪刀、镊子、骨剪、手术针、缝合线、棉花、竹根、石膏粉、防腐剂等。2材料选择由于获得的鸟有时间差或地域环境影响,制作标本时要选择新鲜的。特别是在热带地区,几小时鸟就会腐败,羽毛脱落,内脏分解。而冷冻的鸟类,要…     

制作鲁鱼种经系统浸制标本的方法

鸟类剥制标本应注意的几个操作环节鸟类剥制标本,其操作要求较高,在操作过程中应注意以下几个环节：１剥制（１）剥制时应特别注意的几个部位：如腰背部荐骨至尾骨端、头部、耳孔处、眼眶周围、两翼以及弹孔周围。这些部位皮肤薄、皮肤与肌肉骨骼之间结缔组织多,连接紧...     

穿山甲标本和甲片的DNA提取及PCR扩增

为验证经处理后的穿山甲(Manis spp.)标本和甲片是否可以用于种间分子鉴定标记的开发及个体识别工作,本文在样品的预处理、消化、提取后纯化等方面对传统提取方法进行了改进,分别从穿山甲剥制标本、干皮标本及甲片中提取总DNA;然后用Cyt b基因扩增通用引物、12S rRNA基因全序列扩增引物、RAPD引物及微卫星引物进行了PCR扩增,并对部分扩增结果进行了序列测定.结果表明,除剥制标本的脚底皮张组织外,其他样品基本都可以提取出DNA.以此为模板的PCR扩增中,2种线粒体基因引物扩增出明显目的条带,RAPD引物扩增出种间特异条带,测序结果可用于种间特异性引物及SCAR引物的开发;微卫星引物在甲片样品中扩增稳定,可用于个体识别工作.     

用蟾蜍或蛙做剥制标本实验

在生物学教材中,鸟类剥制标本的制作是规定的实验,但有些学校,由于经费金缺乏,或因操作过程复杂,而随意取消或改为演示进行这一实验,影响了教学效果。几年来,我们用青蛙、蟾蜍替代鸟类,同样能使学生学会剥制标本的方法,达到预期的目的。     

科学家首次揭示早期鸟类曾有四个翅膀

山东天宇自然博物馆郑晓廷、中国科学院古脊椎动物与古人类研究所周忠和与徐星研究员等通过分析山东天宇自然博物馆的化石,发现11件早期原始鸟类标本后肢上长有羽毛的明显迹象。研究人员对这些早期鸟类后肢羽毛或皮肤结构的进行了仔细观察和对比研究,确认这些鸟类后肢羽毛的存在证实了早期鸟类演化过程中曾存在一个与似鸟恐龙相似的四翼阶段,并且后肢羽翼也具备协助飞翔的气动功     

采用风干方法制作鸟类标本

用剥制方法制作鸟类标本,过程复杂,技术要求较高,填装、整形准于掌握。特别是对一些中小型鸟类,要想做成一个栩栩如生的生态标本就更困难了。针对上述缺点,我们对制作中小型鸟类标本做了一些改进,取得了良好的效果。我们采用自然风干的方法,制作中小型鸟类(也包括蝙蝠等适于此种制作方法的其它动物)标本,不仅省去了剥制过程中,剥皮和填装等较难掌握的环节,而且不失其自然形态,易于整形。现将制作方法介绍如下:一、工具和材料的准备镊子、软毛刷、仪眼、烧杯、钟形罩、滑石粉、福尔马林、高锰酸钾。二、制作前的处理     

一种高效快速的鱼类标本基因组DNA提取方法

以95%酒精保存的黄鳝(M onopterus albus)和斑鳢(Channa maculates)标本为材料,采用先沉降DNA再去除杂质的方法从鱼类标本中提取基因组DNA。基因组DNA的琼脂糖凝胶电泳和紫外分光光度法检测以及PCR扩增结果显示,本方法提取的鱼类基因组DNA的电泳主带清晰明亮;A260/A280值在1.7830-2.0144之间;PCR扩增产物条带清晰明亮,且单一整齐没有拖带,表明本方法可从酒精保存的鱼类标本中提取比较纯净的DNA,能够满足一般分子生物学试验需要。与传统苯酚/氯仿法相比,本方法操作简单快速,避免了苯酚等物质对后续实验的影响,可作为一种常规动物组织DNA提取方法。     

免疫胶体金法提取环境标本中细菌DNA技术*

将抗DNA单克隆抗体标记在胶体金颗粒上制成免疫胶体金试剂,提取标本中DNA,直接用于PCR检测,从而建立一种简单、快速、高效的免疫胶体金方法提取环境标本中的DNA。结果表明:应用免疫胶体金试剂可有效去除环境标本中PCR抑制剂,浓缩模板,提高PCR检测敏感度3～4个数量级。操作步骤简单,无需使用有机溶剂,避免环境污染,吸附了DNA的免疫胶体金可直接用于PCR扩增。研制了免疫胶体金试剂并确定其最佳反应条件,有效提高PCR技术在检测现场环境标本中的敏感性和实用性。     

一种通用的基因组DNA提取方法

目的:探讨一种通厢的基因组DNA提取方法.方法:采用改良的膜法分别从动植物组织、外周血、细菌、细胞等标本提取基因组DNA,DNA样品经紫外吸收、琼脂糖凝胶电泳、PCR扩增和酶切进行榆测.结果:该方法提取的基因组DNA纯度较高,电泳条带清晰,DNA质量能满足下游分子生物学研究的需要.结论:该方法简便快速、适用范围广,是提取基因组DNA的一种有效方法.     

Museum genomics: low-cost and high-accuracy genetic data from historical specimens

Natural history collections are unparalleled repositories of geographical and temporal variation in faunal conditions. Molecular studies offer an opportunity to uncover much of this variation; however, genetic studies of historical museum specimens typically rely on extracting highly degraded and chemically modified DNA samples from skins, skulls or other dried samples. Despite this limitation, obtaining short fragments of DNA sequences using traditional PCR amplification of DNA has been the primary method for genetic study of historical specimens. Few laboratories have succeeded in obtaining genome-scale sequences from historical specimens and then only with considerable effort and cost. Here, we describe a low-cost approach using high-throughput next-generation sequencing to obtain reliable genome-scale sequence data from a traditionally preserved mammal skin and skull using a simple extraction protocol. We show that single-nucleotide polymorphisms (SNPs) from the genome sequences obtained independently from the skin and from the skull are highly repeatable compared to a reference genome.     

一种经济高效提取实蝇基因组DNA的方法

单头实蝇高质量基因组DNA的获得为实蝇分子生态学研究奠定了重要基础。本文提出一种经济高效的实蝇基因组DNA提取方法,该方法广泛适用于不同虫态、不同保存条件的实蝇标本,与传统的CTAB法和SDS法相比操作简单、耗时短,得率高。     

DNA microextraction from dried blood spots on filter paper blotters: potential applications to newborn screening

Summary Microextraction of DNA from dried blood specimens would ease specimen transport to centralized laboratory facilities for recombinant DNA diagnosis in the same manner as use of dried blood spots allowed the broad application of screening tests to newborn populations. A method is described which reproducibly yields 0.5g DNA from the dried equivalent of 50l whole blood. Though DNA yields decreased with storage of dried specimens at room temperature, good-quality DNA was still obtained. Sufficient DNA was routinely obtained for Southern blot analysis using repetitive and unique sequences. This microextraction procedure will allow immediate application of molecular genetic technology to direct newborn screening follow-up of disorders amenable to DNA diagnosis, such as sickle cell anemia, and may eventually permit primary DNA screening for specific mutations.     

Rapid and reliable high-throughput methods of DNA extraction for use in barcoding and molecular systematics of mushrooms

We present two methods for DNA extraction from fresh and dried mushrooms that are adaptable to high-throughput sequencing initiatives, such as DNA barcoding. Our results show that these protocols yield ∼85% sequencing success from recently collected materials. Tests with both recent (<2 year) and older (>100 years) specimens reveal that older collections have low success rates and may be an inefficient resource for populating a barcode database. However, our method of extracting DNA from herbarium samples using small amount of tissue is reliable and could be used for important historical specimens. The application of these protocols greatly reduces time, and therefore cost, of generating DNA sequences from mushrooms and other fungi vs. traditional extraction methods. The efficiency of these methods illustrates that standardization and streamlining of sample processing should be shifted from the laboratory to the field.     

A simple and reliable method for DNA extraction from bivalve mantle

A simple and reliable method was developed for extracting genomic DNA from preserved mantle tissues of Pacific oyster Crassostrea gigas for reproducible PCR amplification. The method was based on destruction of the tissue using Proteinase K, Chelex 100 resin, detergents, and urea, followed by preferential capturing of genomic DNA with silica particles. Approximately 5 mg of mantle tissue provided a sufficient quality and quantity of DNA for several hundreds of PCR reactions amplifying the hypervariable mitochondrial DNA intergenic spacer, which is a useful genetic marker for population structure analysis of Pacific oyster. The method can be applied for DNA preparation from not only fresh and frozen but also ethanol-preserved mantle tissues, so this rapid and economical method can serve for investigating a large number of bivalve specimens collected in the field and next transported in ethanol at ambient temperature.     

Rapid, one-step DNA extraction for insect pest identification by using DNA barcodes

Early detection of economically important insects is critical to preventing their establishment as serious pests. To accomplish this, tools for rapid and accurate species identification are needed. DNA barcoding, using short DNA sequences as species "genetic identification tags," has already shown large potential as a tool for rapid and accurate detection of economically important insects. DNA extraction is the critical first step in generating DNA barcodes and can be a rate-limiting step in very large barcoding studies. Consequently, a DNA extraction method that is rapid, easy to use, cost-effective, robust enough to cope with range of qualities and quantities of tissue, and can be adapted to robotic systems will provide the best method for high-throughput production of DNA barcodes. We tested the performance of a new commercial kit (prepGEM), which uses a novel, streamlined approach to DNA extraction, and we compared it with two other commercial kits (ChargeSwitch and Aquapure), which differ in their method of DNA extraction. We compared performance of these kits by measuring percentage of polymerase chain reaction (PCR) success and mean PCR product yield across a variety of arthropod taxa, whichincluded freshly collected, ethanol-preserved, and dried specimens of different ages. ChargeSwitch and prepGEM performed equally well, but they outperformed Aquapure. prepGEM was much faster, easier to use, and cheaper than ChargeSwitch, but ChargeSwitch performed slightly better for older (> 5-yr-old) dried insect specimens. Overall, prepGEM may provide a highly streamlined method of DNA extraction for fresh, ethanol-preserved, and young, dried specimens, especially when adapted for high-throughput, robotic systems.     

More than skin and bones: Comparing extraction methods and alternative sources of DNA from avian museum specimens

Next‐generation sequencing has greatly expanded the utility and value of museum collections by revealing specimens as genomic resources. As the field of museum genomics grows, so does the need for extraction methods that maximize DNA yields. For avian museum specimens, the established method of extracting DNA from toe pads works well for most specimens. However, for some specimens, especially those of birds that are very small or very large, toe pads can be a poor source of DNA. In this study, we apply two DNA extraction methods (phenol–chloroform and silica column) to three different sources of DNA (toe pad, skin punch and bone) from 10 historical avian museum specimens. We show that a modified phenol–chloroform protocol yielded significantly more DNA than a silica column protocol (e.g., Qiagen DNeasy Blood & Tissue Kit) across all tissue types. However, extractions using the silica column protocol contained longer fragments on average than those using the phenol–chloroform protocol, probably as a result of loss of small fragments through the silica column. While toe pads yielded more DNA than skin punches and bone fragments, skin punches proved to be a reliable alternative source of DNA and might be especially appealing when toe pad extractions are impractical. Overall, we found that historical bird museum specimens contain substantial amounts of DNA for genomic studies under most extraction scenarios, but that a phenol–chloroform protocol consistently provides the high quantities of DNA required for most current genomic protocols.     

Hi-Plex targeted sequencing is effective using DNA derived from archival dried blood spots

Many genetic epidemiology resources have collected dried blood spots (predominantly as Guthrie Cards) as an economical and efficient means of archiving sources of DNA, conferring great value to genetic screening methods that are compatible with this medium. We applied Hi-Plex to screen the breast cancer predisposition gene PALB2 in 93 Guthrie Card-derived DNA specimens previously characterized for PALB2 genetic variants via DNA derived from lymphoblastoid cell lines, whole blood, and buffy coat. Of the 93 archival Guthrie Card-derived DNAs, 92 (99%) were processed successfully and sequenced using approximately half of a MiSeq run. From these 92 DNAs, all 59 known variants were detected and no false-positive variant calls were yielded. Fully 98.13% of amplicons (5417/5520) were represented within 15-fold of the median coverage (2786 reads), and 99.98% of amplicons (5519/5520) were represented at a depth of 10 read-pairs or greater. With Hi-Plex, we show for the first time that a High-Plex amplicon-based massively parallel sequencing (MPS) system can be applied effectively to DNA prepared from dried blood spot archival specimens and, as such, can dramatically increase the scopes of both method and resource.     

Retrospective Estimation of Genetic Diversity of an Extinct Oriental White Stork (Ciconia boyciana) Population in Japan Using Mounted Specimens and Implications for Reintroduction Programs

The Japanese regional population of the Oriental white stork (Ciconia boyciana) became extinct in 1986. Mitochondrial DNA (mtDNA) D-loop region from 20 mounted specimens preserved at public facilities in Toyooka City, Hyogo Prefecture, Japan and its vicinity (n = 17), the area inhabited by the last of the Japanese population, and samples originating from China (n = 3) which were kept at a zoo was analyzed. After extracting DNA from small pieces of skin from mounted specimens, a 1210-bp region of the mtDNA D-loop region was analyzed. The haplotypes among 11 specimens of storks captured or found dead at Toyooka City just before the population became extinct were completely identical. Four haplotypes observed among the mounted specimens preserved in the vicinity of Toyooka City were differentiated from those of captive storks originating from China and Russia in a previous study. Therefore, the last Japanese population might have been a genetically unique group. However, phylogenetic analysis using the maximum likelihood method showed that haplotypes found in the Japanese regional population were closely related to the Chinese and Russian lineages (sequence difference = 2.1%). One mounted specimen collected in 1935 at Izushi village, in the vicinity of Toyooka City, showed the same haplotype as the captive storks from China, suggesting that genetic flow may have historically occurred between the populations of Japan and the continent. Recently, reintroduction for the Oriental white stork has been planned in Toyooka City. The planning for the recovery of extinct populations should not only involve translocation of species to the range from which it disappeared, but also reconstruction of regional populations while considering the genetic lineage between the extinct and introduced populations.