目的:以α地中海贫血基因诊断所需的m arker为例,探讨一种制备新的DNA m arker的方法。方法:选取相应实验所能检测到的所有阳性标本及阴性标本1份,常规抽提DNA及PCR扩增,将所有标本的PCR产物混合即可制成该实验所需专用DNA m arker。结...目的:以α地中海贫血基因诊断所需的m arker为例,探讨一种制备新的DNA m arker的方法。方法:选取相应实验所能检测到的所有阳性标本及阴性标本1份,常规抽提DNA及PCR扩增,将所有标本的PCR产物混合即可制成该实验所需专用DNA m arker。结果:3份阳性标本扩增后分别获得2000bp、1800bp、1600bp、1300bp条带,分别对应-3α.7、正常、-4α.2、--SEA的位置,混匀后所得专用m arker包含以上条带。结论:新组合的DNA m arker定位清晰,方法简便、经济、易行。展开更多
Chili pepper (Capsicum spp.) fruit color is an important agronomical trait. It has been known that a large deletion in the 5' upstream region of the Ccs gene generates non-red fruit color in pepper, but the accura...Chili pepper (Capsicum spp.) fruit color is an important agronomical trait. It has been known that a large deletion in the 5' upstream region of the Ccs gene generates non-red fruit color in pepper, but the accurate size and position of the deletion and whether all the non-red cultivars had the same large deletion or not were unclarified. In this study, to identify the Ccs upstream large deletion, we carried out diagnostic PCR using six forward primers at 300 - 900 bp intervals in the 5' untranslated region of Ccs with a fixed reverse primer for a yellow fruit pepper “Sonia Gold”. Then it was revealed that 4430 bp from -3234 bp position in upstream region to 1196 bp position in exon was deleted in Ccs of “Sonia Gold”. The allele having this deletion was named ccs-del. Probably this allele is substantially the same as ccs-p1 having 4879 bp deletion reported previously. Based on the sequence determined, we developed a PCR marker to distinguish ccs-del. Genotyping of 16 cultivars of C. annuum showed that 14 had ccs-del and the remaining two had another mutant allele ccs-3. This result indicates that ccs-del is the most common allele and widely shared in non-red fruit cultivars in C. annuum. Genotyping of 16 cultivars of C. chinense clarified that one cultivar each possessed ccs-del and ccs-3. These results indicate that major alleles responsible for non-red fruit color in C. annuum were shared across species throughout interspecific introgression.展开更多
Informative,portable,and efficient DNA markers have the potential to accelerate genetic gain in cotton breeding.Discovery and widespread application of DNA markers to cotton has
Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have be...Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have been identified in various potato species and most of these R genes have been used in potato breeding. The aim of this study was to develop and validate PCR-based assays for the R genes Rpi-blb1, Rpi-blb2, Rpi-blb3 and Rpi-bt1, to distinguish between late blight resistant and late blight susceptible potato progeny in the given breeding background. A total of 100 breeding progeny were screened for the presence of these R genes and tested for resistance against P. infestans mating type A2, genotype US-8 strain, using detached leaf and tuber rot assays. PCR products for the Rpi-blb1 and Rpi-bt1 resistance genes were identified in the resistant progeny but were absent in the susceptible ones;therefore these PCR assays could differentiate between late blight resistant and susceptible plants. Genotypic data from the DNA markers derived from the Rpi-blb1 and Rpi-bt1 genes was found to correlate with the phenotypic data for foliar late blight but not with data for tuber rot. Our results demonstrate that markers derived from these two R genes could be useful for marker-assisted selection (MAS) for foliar late blight resistance in potato breeding programs.展开更多
Rapid and reliable identification of olive plants using DNA markers has been attempted in the past but the selection of polymorphic regions for discrimination at varietal level remained obscure. Recent sequencing of p...Rapid and reliable identification of olive plants using DNA markers has been attempted in the past but the selection of polymorphic regions for discrimination at varietal level remained obscure. Recent sequencing of plastid genome of the olive flaunts high resolution Cp markers for olive DNA fingerprinting. Using this information, we designed a combination of chloroplast markers to amplify genes recruited in photosynthesis, ribosomal and NADH energy metabolism for varietal identification of olive plants. Concatenated DNA sequences of more than 100 unknown and 10 reference plants samples were analyzed using various bioinformatics and phylogenetic tools. Conserved blocks of nucleotide sequences were detected in multiple alignments. Phylogenetic reconstruction differentiated the unknown plants into various clusters with known varieties. Further narrowing down of the samples through UPGMA tree clearly separated the plants into Arbosana, Frantoio and Koroneiki as the major varieties. Multiple alignments of these clusters revealed important variety specific SNPs including G and T nucleotides at specific positions. Sequence identifying at intra cultivar level was more than 98.79% while it dropped to 97%, and even to 96% at inter varietal level. Furthermore, a neighbor net network analysis separated these three clusters, thus validating the results of UPGMA tree. Over all, out of 100 plants samples, 49 plants were identified that fall into 10 varieties including Arbosana, Carolea, Chetoui, Coratina, Domat, Frantoio, Gemlik, Koroneiki,Leccino and Moraiolo. The maximum number of known plants belongs to Frantoio and Gemlik (8 each). The least number of samples was identified from Carolea, Domat and Moraiolo with 2 samples each. However, 51 plants could not be identified, as plants were not clustered with any of reference control. Our results have implications in on-farm conservation of olive germplasm and provision of genuine material for multiplication of authentic varieties. This strategy can be extended to varietal identification of other plant species.展开更多
利用磁珠法对甘蓝、白菜、芥菜、洋葱、黄瓜、番茄、辣椒等7种形状、大小不同的蔬菜单粒种子和幼苗进行DNA提取,采用内参基因Actin进行实时荧光定量PCR评价DNA质量,利用KASP(kompetitive allele specific PCR)标记检验DNA质量是否达到K...利用磁珠法对甘蓝、白菜、芥菜、洋葱、黄瓜、番茄、辣椒等7种形状、大小不同的蔬菜单粒种子和幼苗进行DNA提取,采用内参基因Actin进行实时荧光定量PCR评价DNA质量,利用KASP(kompetitive allele specific PCR)标记检验DNA质量是否达到KASP检测要求。结果显示,单粒种子磁珠法提取的各蔬菜DNA平均Ct值均处于19~24范围内。KASP检测进一步证实种子提取DNA的质量达到KASP标记分型的要求。综上所述,本研究建立了一种简单、经济、实用性强的单粒种子DNA提取方法,该方法对不同大小的蔬菜种子具有较好的稳定性,且大幅缩短了获得基因型数据的时间。展开更多
对比分析乙肝病毒DNA(HBV-DNA)定量检测与免疫学标志物检测的检测效果。采用随机数字表法从贵州省惠水县人民医院2021年5月—2022年4月收治的乙肝患者中抽取120例进行研究,患者均接受HBV-DNA定量检测和免疫学标志物检测,根据检测结果将...对比分析乙肝病毒DNA(HBV-DNA)定量检测与免疫学标志物检测的检测效果。采用随机数字表法从贵州省惠水县人民医院2021年5月—2022年4月收治的乙肝患者中抽取120例进行研究,患者均接受HBV-DNA定量检测和免疫学标志物检测,根据检测结果将患者分为其他模型组(40例)、小三阳组(40例)、大三阳组(40例)。结果显示,HBV-DNA定量检测:大三阳组高于小三阳组、其他模型组,小三阳组高于其他模型组,组间数据差异具有统计学意义(P<0.05);HBV-DNA检测与免疫学标志物检测阳性率对比:HBV-DNA阳性标本中,HBsAg检出率为96.55%(56/58),HBeAg检出率为48.28%(28/58)。研究发现,临床诊断乙肝病毒(hepatitis B virus,HBV)感染时可采用HBV-DNA定量检测与免疫学标志物检测,这两种检测方式联合检测对于乙肝患者的临床诊治和预后诊断具有重要的参考价值。展开更多
目的研究探讨乙肝孕妇大三阳和小三阳状态中乙型肝炎病毒(Hepatitis B virus,HBV)DNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能水平。方法选取临清市人民医院于2020年1月—2023年1月收治的120例乙肝孕妇作为...目的研究探讨乙肝孕妇大三阳和小三阳状态中乙型肝炎病毒(Hepatitis B virus,HBV)DNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能水平。方法选取临清市人民医院于2020年1月—2023年1月收治的120例乙肝孕妇作为研究对象,根据乙肝两对半检测结果将其分为两组,即大三阳组(n=60)和小三阳组(n=60)。比较两组HBVDNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能指标、肝纤维化标志物含量。结果大三阳组丙氨酸转移酶水平(29.00±7.62)U/L、HBVDNA水平(5.18±1.16)×10^(4)U/mL均高于小三阳组的(26.00±2.85)U/L、(2.08±1.11)×10^(4)U/mL,差异有统计学意义(t=2.856、14.956,P均<0.05)。大三阳组的重组人几丁质酶3样蛋白1水平低于小三阳组,差异有统计学意义(P<0.05)。伴随着HBVDNA载量上升,两组患者甲胎蛋白水平随之增加,但是在同等HBADNA载量下,两组甲胎蛋白水平比较,差异无统计学意义(P>0.05)。结论开展孕前抗乙肝病毒诊疗、孕期风险系数评估,采取科学有效的干预措施,均能够有效抑制乙肝在母婴中的传播。展开更多
目的分析失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与乙型肝炎病毒标志物、生化及凝血等指标的相关性。方法回顾性分析2018年1月至2020年12月我院收治的645例失代偿期乙型肝炎肝硬化患者的临床资料,根据HBV-DNA水平分为4组:阴性组、...目的分析失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与乙型肝炎病毒标志物、生化及凝血等指标的相关性。方法回顾性分析2018年1月至2020年12月我院收治的645例失代偿期乙型肝炎肝硬化患者的临床资料,根据HBV-DNA水平分为4组:阴性组、低病毒载量组(<2.0×10^(3)IU/mL)、中病毒载量组(2.0×10^(3)~2.0×10^(5)IU/mL)和高病毒载量组(>105IU/mL),测定所有患者乙型肝炎病毒血清学标志物(HBV-M)、肝功能相关生化指标(ALT、AST、TBIL、DBIL、ALB)、凝血相关指标(PT、INR、APTT、TT、D-D、PLT),比较4组患者各项指标的差异,并分析各项指标与HBV-DNA水平的相关性。结果在失代偿期乙型肝炎肝硬化患者中,HBsAg和HBeAg的定量数值随着HBV-DNA水平的升高而增高,HBsAg的明显升高主要体现在HBV-DNA阴性组和低病毒载量组之间(779.40 vs 5773.00IU/mL),而HBeAg的表达在中病毒载量组(0.19 COI)和高病毒载量组(7.96 COI)中才出现显著升高,低病毒载量组的HBeAg阳性表达率较低。随着病毒载量的升高,患者的ALT、AST、TBIL、DBIL、D/T、AFP指标均有所升高,而ALB的水平降低;除TT存在显著性差异外,PT、APTT、INR、Fib、D-D、PLT这6个指标差异均无统计学意义。经Pearson相关性分析检验,失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与ALT、AST、TBIL、DBIL、AFP、PT、APTT、TT、D-D、HBsAg、HBeAg水平呈正相关(r>0,P均<0.05),与ALB、Fib水平呈负相关(r<0,P均<0.05),与PLT、HBeAb水平不存在相关性。结论在失代偿期乙型肝炎肝硬化患者中,血清HBV-DNA载量与HBsAg、HBeAg表达水平密切相关,与ALT、AST、TBIL、DBIL、D/T、ALB等肝功能相关生化指标相关,与凝血相关指标存在关联,但在不同HBV-DNA载量组中无差异。HBV-DNA载量可作为HBeAg阴性患者肝脏损害程度的有效预测指标。展开更多
Marker-assisted selection is an important tool in squash (<i>Cucurbita</i> species) breeding. A seed-based genotyping system would not only allow selection of desirable individuals prior to planting, but a...Marker-assisted selection is an important tool in squash (<i>Cucurbita</i> species) breeding. A seed-based genotyping system would not only allow selection of desirable individuals prior to planting, but also reduce the cost associated with leaf-derived DNA genotyping, such as the need for greenhouse facilities and ultra-low-temperature storage freezers. A robust seed-based genotyping system requires a non-destructive sampling method and DNA of sufficient quantity and quality for marker-assisted selection. In the current study, six cultivars representing <i>Cucurbita</i> <i>pepo</i> (Black Beauty and Yellow Crookneck), <i>C</i>. <i>moschata</i> (Butterbush and Fairytale), and <i>C</i>. <i>maxima</i> (Buttercup and Big <span>Max) were used to develop a suitable seed-based genotyping system for squash.</span> Seed chips for DNA extraction were sampled by removing </span><span style="font-family:"">1/3</span><span style="font-family:""> of the distal end, while the remnant seed-embryos were sowed to assess germination potential. <span>Four extraction methods including two column-based commercial kits (CTAB</span> and ENZA) and two detergent-based conventional methods (CTAB and SDS) were assessed for DNA quality and quantity. Utility of extracted DNA for downstream applications was tested by genotyping with SSR and SNP markers. There was no significant difference in germination percentage between whole and cut seeds across the six cultivars. The average DNA concentration across methods ranged from 11.6 ng/μL to 62.6 ng/μl, while the DNA quality (A<sub>260/280</sub>) ranged from 0.89 to 1.95. Although DNA was obtained for all the extraction methods, only EZNA and Favorgen methods yielded DNA of sufficient quality for marker-assisted selection.展开更多
文摘目的:以α地中海贫血基因诊断所需的m arker为例,探讨一种制备新的DNA m arker的方法。方法:选取相应实验所能检测到的所有阳性标本及阴性标本1份,常规抽提DNA及PCR扩增,将所有标本的PCR产物混合即可制成该实验所需专用DNA m arker。结果:3份阳性标本扩增后分别获得2000bp、1800bp、1600bp、1300bp条带,分别对应-3α.7、正常、-4α.2、--SEA的位置,混匀后所得专用m arker包含以上条带。结论:新组合的DNA m arker定位清晰,方法简便、经济、易行。
文摘Chili pepper (Capsicum spp.) fruit color is an important agronomical trait. It has been known that a large deletion in the 5' upstream region of the Ccs gene generates non-red fruit color in pepper, but the accurate size and position of the deletion and whether all the non-red cultivars had the same large deletion or not were unclarified. In this study, to identify the Ccs upstream large deletion, we carried out diagnostic PCR using six forward primers at 300 - 900 bp intervals in the 5' untranslated region of Ccs with a fixed reverse primer for a yellow fruit pepper “Sonia Gold”. Then it was revealed that 4430 bp from -3234 bp position in upstream region to 1196 bp position in exon was deleted in Ccs of “Sonia Gold”. The allele having this deletion was named ccs-del. Probably this allele is substantially the same as ccs-p1 having 4879 bp deletion reported previously. Based on the sequence determined, we developed a PCR marker to distinguish ccs-del. Genotyping of 16 cultivars of C. annuum showed that 14 had ccs-del and the remaining two had another mutant allele ccs-3. This result indicates that ccs-del is the most common allele and widely shared in non-red fruit cultivars in C. annuum. Genotyping of 16 cultivars of C. chinense clarified that one cultivar each possessed ccs-del and ccs-3. These results indicate that major alleles responsible for non-red fruit color in C. annuum were shared across species throughout interspecific introgression.
文摘Informative,portable,and efficient DNA markers have the potential to accelerate genetic gain in cotton breeding.Discovery and widespread application of DNA markers to cotton has
文摘Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have been identified in various potato species and most of these R genes have been used in potato breeding. The aim of this study was to develop and validate PCR-based assays for the R genes Rpi-blb1, Rpi-blb2, Rpi-blb3 and Rpi-bt1, to distinguish between late blight resistant and late blight susceptible potato progeny in the given breeding background. A total of 100 breeding progeny were screened for the presence of these R genes and tested for resistance against P. infestans mating type A2, genotype US-8 strain, using detached leaf and tuber rot assays. PCR products for the Rpi-blb1 and Rpi-bt1 resistance genes were identified in the resistant progeny but were absent in the susceptible ones;therefore these PCR assays could differentiate between late blight resistant and susceptible plants. Genotypic data from the DNA markers derived from the Rpi-blb1 and Rpi-bt1 genes was found to correlate with the phenotypic data for foliar late blight but not with data for tuber rot. Our results demonstrate that markers derived from these two R genes could be useful for marker-assisted selection (MAS) for foliar late blight resistance in potato breeding programs.
文摘Rapid and reliable identification of olive plants using DNA markers has been attempted in the past but the selection of polymorphic regions for discrimination at varietal level remained obscure. Recent sequencing of plastid genome of the olive flaunts high resolution Cp markers for olive DNA fingerprinting. Using this information, we designed a combination of chloroplast markers to amplify genes recruited in photosynthesis, ribosomal and NADH energy metabolism for varietal identification of olive plants. Concatenated DNA sequences of more than 100 unknown and 10 reference plants samples were analyzed using various bioinformatics and phylogenetic tools. Conserved blocks of nucleotide sequences were detected in multiple alignments. Phylogenetic reconstruction differentiated the unknown plants into various clusters with known varieties. Further narrowing down of the samples through UPGMA tree clearly separated the plants into Arbosana, Frantoio and Koroneiki as the major varieties. Multiple alignments of these clusters revealed important variety specific SNPs including G and T nucleotides at specific positions. Sequence identifying at intra cultivar level was more than 98.79% while it dropped to 97%, and even to 96% at inter varietal level. Furthermore, a neighbor net network analysis separated these three clusters, thus validating the results of UPGMA tree. Over all, out of 100 plants samples, 49 plants were identified that fall into 10 varieties including Arbosana, Carolea, Chetoui, Coratina, Domat, Frantoio, Gemlik, Koroneiki,Leccino and Moraiolo. The maximum number of known plants belongs to Frantoio and Gemlik (8 each). The least number of samples was identified from Carolea, Domat and Moraiolo with 2 samples each. However, 51 plants could not be identified, as plants were not clustered with any of reference control. Our results have implications in on-farm conservation of olive germplasm and provision of genuine material for multiplication of authentic varieties. This strategy can be extended to varietal identification of other plant species.
文摘利用磁珠法对甘蓝、白菜、芥菜、洋葱、黄瓜、番茄、辣椒等7种形状、大小不同的蔬菜单粒种子和幼苗进行DNA提取,采用内参基因Actin进行实时荧光定量PCR评价DNA质量,利用KASP(kompetitive allele specific PCR)标记检验DNA质量是否达到KASP检测要求。结果显示,单粒种子磁珠法提取的各蔬菜DNA平均Ct值均处于19~24范围内。KASP检测进一步证实种子提取DNA的质量达到KASP标记分型的要求。综上所述,本研究建立了一种简单、经济、实用性强的单粒种子DNA提取方法,该方法对不同大小的蔬菜种子具有较好的稳定性,且大幅缩短了获得基因型数据的时间。
文摘对比分析乙肝病毒DNA(HBV-DNA)定量检测与免疫学标志物检测的检测效果。采用随机数字表法从贵州省惠水县人民医院2021年5月—2022年4月收治的乙肝患者中抽取120例进行研究,患者均接受HBV-DNA定量检测和免疫学标志物检测,根据检测结果将患者分为其他模型组(40例)、小三阳组(40例)、大三阳组(40例)。结果显示,HBV-DNA定量检测:大三阳组高于小三阳组、其他模型组,小三阳组高于其他模型组,组间数据差异具有统计学意义(P<0.05);HBV-DNA检测与免疫学标志物检测阳性率对比:HBV-DNA阳性标本中,HBsAg检出率为96.55%(56/58),HBeAg检出率为48.28%(28/58)。研究发现,临床诊断乙肝病毒(hepatitis B virus,HBV)感染时可采用HBV-DNA定量检测与免疫学标志物检测,这两种检测方式联合检测对于乙肝患者的临床诊治和预后诊断具有重要的参考价值。
文摘目的研究探讨乙肝孕妇大三阳和小三阳状态中乙型肝炎病毒(Hepatitis B virus,HBV)DNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能水平。方法选取临清市人民医院于2020年1月—2023年1月收治的120例乙肝孕妇作为研究对象,根据乙肝两对半检测结果将其分为两组,即大三阳组(n=60)和小三阳组(n=60)。比较两组HBVDNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能指标、肝纤维化标志物含量。结果大三阳组丙氨酸转移酶水平(29.00±7.62)U/L、HBVDNA水平(5.18±1.16)×10^(4)U/mL均高于小三阳组的(26.00±2.85)U/L、(2.08±1.11)×10^(4)U/mL,差异有统计学意义(t=2.856、14.956,P均<0.05)。大三阳组的重组人几丁质酶3样蛋白1水平低于小三阳组,差异有统计学意义(P<0.05)。伴随着HBVDNA载量上升,两组患者甲胎蛋白水平随之增加,但是在同等HBADNA载量下,两组甲胎蛋白水平比较,差异无统计学意义(P>0.05)。结论开展孕前抗乙肝病毒诊疗、孕期风险系数评估,采取科学有效的干预措施,均能够有效抑制乙肝在母婴中的传播。
文摘目的分析失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与乙型肝炎病毒标志物、生化及凝血等指标的相关性。方法回顾性分析2018年1月至2020年12月我院收治的645例失代偿期乙型肝炎肝硬化患者的临床资料,根据HBV-DNA水平分为4组:阴性组、低病毒载量组(<2.0×10^(3)IU/mL)、中病毒载量组(2.0×10^(3)~2.0×10^(5)IU/mL)和高病毒载量组(>105IU/mL),测定所有患者乙型肝炎病毒血清学标志物(HBV-M)、肝功能相关生化指标(ALT、AST、TBIL、DBIL、ALB)、凝血相关指标(PT、INR、APTT、TT、D-D、PLT),比较4组患者各项指标的差异,并分析各项指标与HBV-DNA水平的相关性。结果在失代偿期乙型肝炎肝硬化患者中,HBsAg和HBeAg的定量数值随着HBV-DNA水平的升高而增高,HBsAg的明显升高主要体现在HBV-DNA阴性组和低病毒载量组之间(779.40 vs 5773.00IU/mL),而HBeAg的表达在中病毒载量组(0.19 COI)和高病毒载量组(7.96 COI)中才出现显著升高,低病毒载量组的HBeAg阳性表达率较低。随着病毒载量的升高,患者的ALT、AST、TBIL、DBIL、D/T、AFP指标均有所升高,而ALB的水平降低;除TT存在显著性差异外,PT、APTT、INR、Fib、D-D、PLT这6个指标差异均无统计学意义。经Pearson相关性分析检验,失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与ALT、AST、TBIL、DBIL、AFP、PT、APTT、TT、D-D、HBsAg、HBeAg水平呈正相关(r>0,P均<0.05),与ALB、Fib水平呈负相关(r<0,P均<0.05),与PLT、HBeAb水平不存在相关性。结论在失代偿期乙型肝炎肝硬化患者中,血清HBV-DNA载量与HBsAg、HBeAg表达水平密切相关,与ALT、AST、TBIL、DBIL、D/T、ALB等肝功能相关生化指标相关,与凝血相关指标存在关联,但在不同HBV-DNA载量组中无差异。HBV-DNA载量可作为HBeAg阴性患者肝脏损害程度的有效预测指标。
文摘Marker-assisted selection is an important tool in squash (<i>Cucurbita</i> species) breeding. A seed-based genotyping system would not only allow selection of desirable individuals prior to planting, but also reduce the cost associated with leaf-derived DNA genotyping, such as the need for greenhouse facilities and ultra-low-temperature storage freezers. A robust seed-based genotyping system requires a non-destructive sampling method and DNA of sufficient quantity and quality for marker-assisted selection. In the current study, six cultivars representing <i>Cucurbita</i> <i>pepo</i> (Black Beauty and Yellow Crookneck), <i>C</i>. <i>moschata</i> (Butterbush and Fairytale), and <i>C</i>. <i>maxima</i> (Buttercup and Big <span>Max) were used to develop a suitable seed-based genotyping system for squash.</span> Seed chips for DNA extraction were sampled by removing </span><span style="font-family:"">1/3</span><span style="font-family:""> of the distal end, while the remnant seed-embryos were sowed to assess germination potential. <span>Four extraction methods including two column-based commercial kits (CTAB</span> and ENZA) and two detergent-based conventional methods (CTAB and SDS) were assessed for DNA quality and quantity. Utility of extracted DNA for downstream applications was tested by genotyping with SSR and SNP markers. There was no significant difference in germination percentage between whole and cut seeds across the six cultivars. The average DNA concentration across methods ranged from 11.6 ng/μL to 62.6 ng/μl, while the DNA quality (A<sub>260/280</sub>) ranged from 0.89 to 1.95. Although DNA was obtained for all the extraction methods, only EZNA and Favorgen methods yielded DNA of sufficient quality for marker-assisted selection.
