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Cadmium chloride down-regulates the expression of Rad51 in HC11 cells and reduces knock-in efficiency

  • Ga-Yeon Kim;Man-Jong Kang
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.3
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    • pp.99-108
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    • 2023
  • Background: Efficient gene editing technology is needed for successful knock-in. Homologous recombination (HR) is a major double-strand break repair pathway that can be utilized for accurately inserting foreign genes into the genome. HR occurs during the S/G2 phase, and the DNA mismatch repair (MMR) pathway is inextricably linked to HR to maintain HR fidelity. This study was conducted to investigate the effect of inhibiting MMR-related genes using CdCl2, an MMR-related gene inhibitor, on HR efficiency in HC11 cells. Methods: The mRNA and protein expression levels of MMR-related genes (Msh2, Msh3, Msh6, Mlh1, Pms2), the HR-related gene Rad51, and the NHEJ-related gene DNA Ligase IV were assessed in HC11 cells treated with 10 μM of CdCl2 for 48 hours. In addition, HC11 cells were transfected with a CRISPR/sgRNA expression vector and a knock-in vector targeting Exon3 of the mouse-beta casein locus, and treated with 10 μM cadmium for 48 hours. The knock-in efficiency was monitored through PCR. Results: The treatment of HC11 cells with a high-dose of CdCl2 decreased the mRNA expression of the HR-related gene Rad51 in HC11 cells. In addition, the inhibition of MMR-related genes through CdCl2 treatment did not lead to an increase in knock-in efficiency. Conclusions: The inhibition of MMR-related gene expression through high-dose CdCl2 treatment reduces the expression of the HR-related gene Rad51, which is active during recombination. Therefore, it was determined that CdCl2 is an inappropriate compound for improving HR efficiency.

Properties of Cu Pillar Bump Joints during Isothermal Aging (등온 시효 처리에 따른 Cu Pillar Bump 접합부 특성)

  • Eun-Su Jang;Eun-Chae Noh;So-Jeong Na;Jeong-Won Yoon
    • Journal of the Microelectronics and Packaging Society
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    • v.31 no.1
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    • pp.35-42
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    • 2024
  • Recently, with the miniaturization and high integration of semiconductor chips, the bump bridge phenomenon caused by fine pitches is drawing attention as a problem. Accordingly, Cu pillar bump, which can minimize the bump bridge phenomenon, is widely applied in the semiconductor package industry for fine pitch applications. When exposed to a high-temperature environment, the thickness of the intermetallic compound (IMC) formed at the joint interface increases, and at the same time, Kirkendall void is formed and grown inside some IMC/Cu and IMC interfaces. Therefore, it is important to control the excessive growth of IMC and the formation and growth of Kirkendall voids because they weaken the mechanical reliability of the joints. Therefore, in this study, isothermal aging evaluation of Cu pillar bump joints with a CS (Cu+ Sn-1.8Ag Solder) structure was performed and the corresponding results was reported.

Phylogenetic Relationships of Jeju Dogs to Other Domestic and Foreign Dog Breeds Determined by Using mtDNA D-loop Sequences (mtDNA D-loop의 염기서열에 의한 제주견과 우리나라 재래견 및 외국견품종과의 유연관계)

  • Kim, Mi-Gyoung;Kim, Nam-Young;Lee, Sung-Soo;Kim, Ky-IL;Yang, Young-Hoon
    • Journal of Animal Science and Technology
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    • v.53 no.4
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    • pp.303-310
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    • 2011
  • Phylogenetic relationships of Jeju dogs to other domestic and foreign dog breeds were assessed using mtDNA D-loop sequences. Neighbor-joining trees were constructed using complete sequences (970 bp excluding the tandem repeat region) determined for five Cheju, four Jindo, four Sapsaree, five Pungsan, two of each East and West Laika dogs (Canis familiaris), two gray wolves (Canis lupus) and two coyotes (Canis latrans) and also published complete sequences for dogs. Coyote sequences were used as outgroups. In addition, a total of 214 haplotypes of 598bp D-loop sequences from 30 dog breeds were collected from GenBank and used to investigate genetic structure of population. In the analyses of full D-loop sequence variation and the phylogenetic trees constructed by neighbor-joining method, neither haplotypes nor clades specific for any domestic dog breeds were observed. The inter-species sequence variation (4.51%) between domestic dogs and wolves was much higher than the intra-species sequence variation within domestic dogs (1.63%) and wolves (3.64%). The divergence of the dog and wolf occurred approximately 1~2 million years ago based on these values. The taxa of Jeju dog breed in the phylogenetic tree are clustered separately and intermingled with other taxa of breeds, suggesting that active crossbreeding of Jeju dogs with other domestic breeds.

A Study on Recognition Methodology and Deduction Improvement Factors of the Registration Process for the Efficient Use of National Research Facilities & Equipments (국가연구시설.장비의 효율적 활용을 위한 인식조사와 등록프로세스 개선요인 도출)

  • Yum, DongKi;Shin, JinGyu
    • Journal of Korea Technology Innovation Society
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    • v.17 no.4
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    • pp.733-762
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    • 2014
  • The government mandates that national research facilities & equipments through R&D business budget should be registered on the National Science and Technology Information Service (NTIS) for the purpose of the efficient use of the research facilities & equipments. This study is to contribute to the national policies on the efficient management of the research facilities & equipments by recognition methodology with the university's members and analysis of the impact factors of the universities' registration process improvement through the Define level and Measure level of the Six Sigma DAMIC. The survey and interview were conducted on research directors, professors joining university administration, graduate students, researchers, and staffs of A University. The findings are the lack of understanding specific steps and life-cycle management of research facilities & equipments. It is necessary to collect suggestions from universities and pursue policies considered the unique characteristics of the university for advanced operating and maximizing use of university's national research facilities & equipments. Research facilities & equipments enrollment compliance rate and registration accuracy were selected as CTQ-Y through the Six Sigma. 72 potential cause variables were derived through Process Map and C & E Diagram. 13 variables were determined as core potential factors through the X-Y Matrix and Pareto Chart. Research institutions should maximize utilization of research facilities & equipments through deriving a potential variables of the process improvements and designing a detail improvements based on the characteristics of each institutions.

Temperature Measurement of Flip Chip Joints with Peripheral Array of Solder Bumps (페리퍼럴어레이 플립칩의 온도 분포 특성)

  • Cho Bon-Goo;Lee Taek-Yeong;Lee Jongwon;Kim Jun-Ki;Kim Gangbeom
    • Journal of the Microelectronics and Packaging Society
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    • v.12 no.3 s.36
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    • pp.243-251
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    • 2005
  • The distribution of temperature of flip chipped device with peripheral solder bump array was measured with variables, such as the locations and geometries of heater, the size of device, the size of passivation opening. The highest temperature was measured with the larger device, $3.0(mm)\times3.0(mm)$, which has the smallest heater at the center of device and the circular passivation opening. For 2 (watts) power input, the device shows the highest temperature of about $110(^{\circ}C)$. In contrast, the smaller device, $1.5(mm)\times1.8(mm)$, shows that of $90(^{\circ}C)$. In addition to the size effect, the increase of passivation opening size decreased the maximum temperature by about $10(^{\circ}C)$. From the measurement, the temperature of device could be controlled with the size and geometry of heater, the size of device and the size and geometry of passivation opening.

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Mechanical Properties and Microstructural Analysis of Sn-40Bi-X Alloys (Sn-40Bi-X 합금의 기계적 물성과 미세조직 분석)

  • Lee, Jong-Hyun;Kim, Ju-Hyung;Hyun, Chang-Yong
    • Proceedings of the KWS Conference
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    • 2010.05a
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    • pp.79-79
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    • 2010
  • 저온용 무연 솔더의 대표 조성으로 고려되고 있는 Sn-58Bi(융점: $138^{\circ}C$) 공정(eutectic) 조성은 우수한 강도에도 불구하고 연성(ductility) 측면에서의 문제점이 지속적으로 보고되고 있다. 따라서 이 합금계의 연성을 최대로 개선시킬 수 있으면서도 실제 상용화가 가능한 합금 조성의 개발 연구가 요청된다. 본 연구에서는 Sn-Bi 2원계 조성에서 최대의 연성을 나타내는 것으로 보고된 Sn-40Bi 조성에 미량의 합금원소를 첨가함으로써 최대의 연성을 확보하는 한편, 그 연성 특성이 변형속도에 어느 정도 민감한지를 인장 실험을 통해 결정하고자 하였다. 합금원소로는 0.1~0.5 wt%의 Ag, Mn, In, Cu를 선택하였으며, 인장 시편을 제조하여 $10^{-2}$, $10^{-3}$, $10^{-4}\;s^{-1}$의 3종류로 변형속도를 변형시켜가며 응력-변형 곡선(stress-strain curve)을 측정하였고, 조성별, 변형속도별로 최대인장강도(ultimate tensile stress, UTS) 및 연신율 결과들을 정리하였다. 합금원소를 첨가한 조성의 경우는 모든 시험 조건에서 Sn-40Bi보다 우수한 연신률을 나타내는 것으로 측정되었으나, $10^{-2}\;s^{-1}$의 빠른 변형속도에서는 그 향상 정도가 상대적으로 감소하는 경향이 관찰되었다. 특히 Sn-40Bi-0.5Ag 조성의 경우 느린 변형속도에서 특히 눈에 띄는 연신률 값을 나타내며, 모든 변형속도 조건에서 가장 우수한 연성을 나타내었다. 한편 Sn-40Bi-0.1Cu 조성의 경우 변형속도에 따른 연신률의 변화 정도, 즉, 변형속도에 따른 연신률의 민감도가 매우 커 $10^{-4}\;s^{-1}$ 속도에서는 Sn-40Bi-0.5Ag에 버금가는 연신률 값이 측정되었으나, $10^{-2}\;s^{-1}$ 속도에서는 가장 나쁜 연신률 특성을 보여주었다. Sn-40Bi-0.2Mn 조성은 최고의 연신률 향상 특성을 나타내지는 않았으나, In을 첨가한 경우보다는 대체적으로 우수한 연성을 나타내었다. 이상의 각 합금별 연성 특성은 인장시험 전의 미세조직 관찰 결과와 인장시험 후 파면부의 조직변화 관찰 결과로부터 해석되었다. 그 결과 석출상의 형성 여부, 인장 시험 중 재결정 조직의 형성 여부, 라멜라(lamellar) 조직의 분율과 라멜라 간격(lamellar spacing)의 정도 또는 $\beta$-Sn과 라멜라 조직 사이의 결정립계와 라멜라 조직 내 결정립계에서의 슬라이딩 모드(sliding mode) 변형 정도, 석출상의 크기와 분포 정도 등이 연신률 및 변형속도 민감도와 같은 연성 특성에 가장 큰 영향을 미치는 인자인 것으로 분석되었다.

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Fault Tolerance for IEEE 1588 Based on Network Bonding (네트워크 본딩 기술을 기반한 IEEE 1588의 고장 허용 기술 연구)

  • Altaha, Mustafa;Rhee, Jong Myung
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.11 no.4
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    • pp.331-339
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    • 2018
  • The IEEE 1588, commonly known as a precision time protocol (PTP), is a standard for precise clock synchronization that maintains networked measurements and control systems. The best master clock (BMC) algorithm is currently used to establish the master-slave hierarchy for PTP. The BMC allows a slave clock to automatically take over the duties of the master when the slave is disconnected due to a link failure and loses its synchronization; the slave clock depends on a timer to compensate for the failure of the master. However, the BMC algorithm does not provide a fast recovery mechanism in the case of a master failure. In this paper, we propose a technique that combines the IEEE 1588 with network bonding to provide a faster recovery mechanism in the case of a master failure. This technique is implemented by utilizing a pre-existing library PTP daemon (Ptpd) in Linux system, with a specific profile of the IEEE 1588 and it's controlled through bonding modes. Network bonding is a process of combining or joining two or more network interfaces together into a single interface. Network bonding offers performance improvements and redundancy. If one link fails, the other link will work immediately. It can be used in situations where fault tolerance, redundancy, or load balancing networks are needed. The results show combining IEEE 1588 with network bonding enables an incredible shorter recovery time than simply just relying on the IEEE 1588 recovery method alone.

Evaluation of High-Temperature Tensile Property of Diffusion Bond of Austenitic Alloys for S-CO2 Cycle Heat Exchangers (고온 S-CO2 사이클 열교환기용 스테인리스강 및 Fe-Cr-Ni 합금 확산 접합부의 고온 인장 특성평가)

  • Hong, Sunghoon;Sah, Injin;Jang, Changheui
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.38 no.12
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    • pp.1421-1426
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    • 2014
  • To improve the inherent safety of the sodium-cooled fast reactor (SFR), the supercritical $CO_2$ ($S-CO_2$) Brayton cycle is being considered as an alternative power conversion system to steam the Rankine cycle. In the $S-CO_2$ system, a PCHE (printed circuit heat exchanger) is being considered. In this type of heat exchangers, diffusion bonding is used for joining the thin plates. In this study, the diffusion bonding characteristics of various austenitic alloys were evaluated. The tensile properties were measured at temperatures starting from the room temperature up to $650^{\circ}C$. For the 316H and 347H types of stainless steel, the tensile ductility was well maintained up to $550^{\circ}C$. However, the Incoloy 800HT showed lower strength and ductility at all temperatures. The microstructure near the bond line was examined to understand the reason for the loss of ductility at high temperatures.

Cold Crack Susceptibility of 700 MPa welding Consumable According Microstructure (700MPa급 용착금속의 미세조직에 따른 저온균열 감수성)

  • Seo, Jun-Seok;Kim, H.J.;Ryoo, H.S.;Park, C.K.;Lee, C.H.
    • Proceedings of the KWS Conference
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    • 2009.11a
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    • pp.46-46
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    • 2009
  • 과거 고강도강 용접부에서 발생하는 저온균열은 주로 용접열영향부에서 발생하였는데, 이러한 문제점을 해결하기 위하여 강재 메이커들은 고강도강의 용접성을 향상시키고자 노력하였다. 이러한 노력의 결과로 TMCP, HSLA 강 등이 개발되었고 이들 강재는 예열온도를 저하시킬 수 있다는 장점 때문에 보편화되어 사용되었다. 이러한 강재는 모재 예열온도를 기준으로 적용하게 되면 용착금속에서 저온균열이 발생하는 경우가 있다. 따라서 이제는 용접재료의 용접성, 즉 용접재료의 저온균열 저항성을 평가 할 수 있는 기법이 요구된다. 본 연구의 목적은 용착금속의 저온균열 저항성을 평가하는 것인데, 저온균열 저항성은 용착금속의 미세조직에 따라 다르게 나타날 수 있다. 용착금속의 합금조성은 기본적으로 용착금속에 요구되는 최저 강도와 충격인성을 만족할 수 있도록 설계한다. 하지만 유사한 강도의 유사한 합금조성이더라도 일부 합금 성분에 의해 용착금속의 미세조직들은 상이하게 나타날 수 있는데, 미세조직 특성에 의하여 용착금속의 강도와 저온인성이 결정된다. 용착금속의 저온균열 저항성을 평가하기위하여 Gapped Bead-on-Groove(G-BOG) 시험에 사용된 모재는 50mm 두께의 mild steel을 사용하였으며, 모재의 희석을 방지하기위해 15mm 깊이로 V-groove 가공 후 buttering 용접 하였다. 용접된 시편은 다시 5mm 깊이로 V-groove로 2차 가공 후 Ar + 20% $Co_2$ gas를 사용하여 용접하였다. 용접재료는 ER-100S-G grade로 비슷한 합금조성을 갖는 2 종류를 사용하였다. A용접재료는 Ti 이 0.1% 함유 되었으며, B용접재료는 Ti 함유되지 않은 것을 사용하였다. 또한 예열 온도에 따라 저온균열 감수성을 평가하기위하여 모재의 예열온도를 각각 상온, $50^{\circ}C,\;75^{\circ}C,\;100^{\circ}C$로 하여 실험을 진행하였다. 용착금속의 미세조직을 확인해본 결과 Ti 함유된 A 용착금속 미세조직은 대부분 침상형페라이트로 나타났으며, Ti 함유되지 않은 B 용착금속 미세조직은 대부분 베이나이트로 나타났다. G-BOG 시험 결과 Ti 함유된 A 시편이 Ti 함유되지 않은 B 시편보다 저온균열 발생량이 적었다. 이는 용착금속의 미세조직분포 및 특성에 따라 저온균열감수성이 다르다는 것을 나타낸다.

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A Study on Welding Deformation of thin plate block in PCTC (PCTC 박판 블록 용접 변형에 관한 연구)

  • Kang, Serng-Ku;Yang, Jong-Su;Kim, Ho-Kyeong
    • Proceedings of the KWS Conference
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    • 2009.11a
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    • pp.97-97
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    • 2009
  • The use of thin plate increases due to the need for light weight in large ship. Thin plate is easily distorted and has residual stress by welding heat. Therefore, the thin plate should be carefully joined to minimize the welding deformation which costs time and money for repair. For one effort to reduce welding deformation, it is very useful to predict welding deformation before welding execution. There are two methods to analyze welding deformation. One is simple linear analysis. The other is nonlinear analysis. The simple linear analysis is elastic analysis using the equivalent load method or inherent strain method from welding experiments. The nonlinear analysis is thermo-elastic analysis which gives consideration to the nonlinearity of material dependent on temperature and time, welding current, voltage, speed, sequence and constraint. In this study, the welding deformation is analyzed by using thermo-elastic method for PCTC(Pure Car and Truck Carrier) which carries cars and trucks. PCTC uses thin plates of 6mm thickness which is susceptible to welding heat. The analysis dimension is 19,200mm(length) * 13,825mm(width) * 376mm(height). MARC and MENTAT are used as pre and post processor and solver. The boundary conditions are based on the real situation in shipyard. The simulations contain convection and gravity. The material of the thin block is mild steel with $235N/mm^2$ yield strength. Its nonlinearity of conductivity, specific heat, Young's modulus and yield strength is applied in simulations. Welding is done in two pass. First pass lasts 2,100 second, then it rests for 900 second, then second pass lasts 2,100 second and then it rests for 20,000 second. The displacement at 0 sec is caused by its own weight. It is maximum 19mm at the free side. The welding line expands, shrinks during welding and finally experiences shrinkage. It results in angular distortion of thin block. Final maximum displacement, 17mm occurs around welding line. The maximum residual stress happens at the welding line, where the stress is above the yield strength. Also, the maximum equivalent plastic strain occurs at the welding line. The plastic strain of first pass is more than that of second pass. The flatness of plate in longitudinal direction is calculated in parallel with the direction of girder and compared with deformation standard of ${\pm}15mm$. Calculated value is within the standard range. The flatness of plate in transverse direction is calculated in perpendicular to the direction of girder and compared with deformation standard of ${\pm}6mm$. It satisfies the standard. Buckle of plate is calculated between each longitudinal and compared with the deformation standard. All buckle value is within the standard range of ${\pm}6mm$.

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