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ÀüÁ¦Ç°º¸±â
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Cloning °ü·Ã
>
¼ö½ÄÈ¿¼Ò£¯DNA Polymerase¡¢RNA Polymerase
> T4 DNA Polymerase
Cloned
T4 DNA Polymerase
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°¡°Ý
(ºÎ°¡¼¼º°µµ)
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Takara
2040A
T4 DNA Polymerase
100 U
72,000¿ø
90,000¿ø
09.02 ~ 10.31
Takara
2040B
T4 DNA Polymerase
500 U (2040A x 5)
405,000¿ø
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DNA Ligation
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½ÇÇè ¸ñÀûº°·Î ÃßõÇÏ´Â..
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Kilo-Sequence ¿ë Delet..
Mighty Cloning Reagent..
Mighty TA-cloning Kit
Mighty TA-cloning Reag..
TaKaRa LA PCR¢â in vit..
TaKaRa T-Vector pMD20
In-Fusion Cloning
[Overview] In-Fusion¢ç..
[°³¿ä] So Easy Product
Online Tools for In-Fu..
[NEW] In-Fusion¢ç Snap..
[NEW] µ¿°á°ÇÁ¶ In-Fusi..
[Kit] In-Fusion¢ç Clon..
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Genomic DNA ÁõÆø¿ë Kit
GenomeWalkerKit
Vector (Cloning ¿ë)
M13 mp18 Single Strand..
pBR322 DNA
pKF18k-2/19k-3 DNA
pSTV28/29 DNA
pUC118/119 type vector
pUC18/19 type vector
TaKaRa T-Vector pMD20
Competent Cells & Electro-..
Agrobacterium tumefaci..
Chaperone Competent Ce..
E. coli Competent Cell..
E. coli Competent Cell..
E. coli Electro-Cells ..
¼ö½ÄÈ¿¼Ò¿¡ ´ëÇÏ¿©
¼ö½ÄÈ¿¼Ò ¼øµµ À϶÷
¼ö½ÄÈ¿¼Òº° ¿ëµµ
¼ö½ÄÈ¿¼Ò£¯Ligase
E. coli DNA Ligase
T4 DNA Ligase
T4 RNA Ligase
¼ö½ÄÈ¿¼Ò£¯Alkaline Phospha..
Alkaline Phosphatase (..
Alkaline Phosphatase (..
Alkaline Phosphatase (..
T4 Polynucleotide Kina..
¼ö½ÄÈ¿¼Ò£¯DNA Polymerase¡¢..
DNA Polymerase I (E. ..
Klenow Fragment (Large..
PrimeCap¢â T7 RNA Poly..
T4 DNA Polymerase
Terminal Deoxynucleoti..
¼ö½ÄÈ¿¼Ò£¯Nuclease¡¢Topois..
BAL 31 Nuclease
DNA Topoisomerase I
Exonuclease I
Exonuclease III
Micrococcal Nuclease
Mung Bean Nuclease
NucleoGone¢â Endonucle..
Recombinant DNase I (R..
Ribonuclease H (RNase ..
S1 Nuclease
Nucleotide
Deoxyribonucleoside 8..
dNTP Mixture
¹èÁö ¡¤ Buffer (ºÐ¸»)
AccuGENE 20¡¿SSPE Buff..
Bovine Serum Albumin S..
IPTG£¨dioxane free£©£¨..
X-Gal£¨5-Bromo-4-Chlor..
±× ¿Ü (À¯Àü°øÇÐ °ü·Ã)
DNA-OFF¢ç
Proteinase K
Westase (È¿¸ð ¼¼Æ÷º® ..
Yatalase
Yatalase -Plus-
Á¦Ç°¼³¸í
º» È¿¼Ò´Â ÁÖÇü DNA¿Í primer°¡ Á¸ÀçÇÒ ¶§ dNTP¸¦ ±âÁú·Î, ÁÖÇü¿¡ »óº¸ÀûÀÎ DNA¸¦ 5¡¯¡æ3¡¯ ¹æÇâÀ¸·Î ÇÕ¼ºÇÑ´Ù. Klenow Fragment¿¡ ºñÇÏ¿© ¾à 100~1000¹è °ÇÑ ´ÜÀÏ °¡´Ú DNA¿¡ ƯÀÌÀûÀÎ 3¡¯¡æ5¡¯ exonuclease È°¼ºÀÌ Æ÷ÇԵǾî ÀÖÁö¸¸ 5¡¯¡æ3¡¯ exonuclease È°¼ºÀº ¾ø´Ù.
º¸Á¸
-20¡É
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5 U/§¡
Çü»ó
200 mM
Potassium Phosphate ¿ÏÃæ¿ë¾× (pH6.5)
1 mM
DTT
50%
Glycerol
÷ºÎ buffer Á¶¼º (10x)
330 mM
Tris-acetate (pH7.9)
660 mM
CH
3
COOK
100 mM
(CH
3
COO)
2
Mg
5 mM
DTT
0.1%
BSA [º°Ã·]
È°¼ºÀÇ Á¤ÀÇ
37¡É, pH8.8¿¡¼ ¿ º¯¼º ¼Û¾ÆÁö Èä¼± DNA¸¦ ÁÖÇü ¹× primer·Î ÇÏ¿© 30 ºÐ°£ 10 nmolÀÇ ¸ðµç nucleotide¸¦ »ê¼ºÀÇ Ä§Àü¹°·Î(acid-insoluble products) ÇÕ¼ºÇÏ´Â È¿¼ÒÈ°¼ºÀ» 1 UÀ¸·Î ÇÑ´Ù.
È°¼º ÃøÁ¤¿ë ¹ÝÀÀ¾× Á¶¼º
67 mM
Tris-HCl (pH8.8)
6.7 mM
MgCl
2
16.6 mM
(NH4)
2
SO
4
1 mM
DTT
6.7 ¥ìM
EDTA
0.0167%
BSA
0.2 §·/§¢
±âÁú DNA
°¢ 330 ¥ìM
dCTP, dATP, dGTP
330 ¥ìM
[
3
H]GTP
»ç¿ë»óÀÇ ÁÖÀÇ
- È¿¼Ò È°¼ºÀ» À§Çؼ Mg
2+
°¡ ÇÊ¿äÇÏ°í, ÃÖ´ë È°¼ºÀ» À§Çؼ´Â SH ȯ¿øÁ¦¸¦ ÷°¡ÇÑ´Ù.
- ¹ÝÀÀ ÀüüÀÇ ÀÌ¿ÂÀÌ 100 mMÀ» ³ÑÀ» °æ¿ì ¹ÝÀÀÀÌ ÀúÇصȴÙ.
- 10x ÷ºÎ buffer¿¡ 0.1% BSA¸¦ ¹Ù·Î ³ÖÀ» °æ¿ì ´ë·®ÀÇ ¹é»ö ħÀüÀÌ »ý±â¹Ç·Î ¹ÝÀÀ¾×À» Á¶Á¦ÇÒ ¶§ ´ÙÀ½ÀÇ ¼ø¼·Î ½Ã¾àÀ» ÷°¡ÇÑ´Ù.
¸ê±Õ Á¤Á¦¼ö ¡æ 10x ÷ºÎ buffer ¡æ 0.1% BSA ¡æ ±âÁú DNA
¿ëµµ
- DNA ÀýÆíÀÇ ÆòÈ° ¸»´ÜÈ
- 3¡¯¡æ5¡¯ exonuclease È°¼ºÀ» ÀÌ¿ëÇÑ replacement synthesis¿¡ ÀÇÇÑ DNAÀÇ 3¡¯-¸»´Ü Ç¥½Ä
±â¿ø
Escherichia coli
carrying the plasmid containing phage T4 DNA polymerase gene
ÀϹÝÀû ¼ºÁú
- ºÐÀÚ·®: ¾à 114,000
- Subunit
´Ü¼øÇÑ polypeptide
È°¼º¿¡ ÇʼöÀûÀÎ 15°³ÀÇ cysteine±â¸¦ °¡Áö°í ÀÖ´Ù.
- º¸Á¶ÀÎÀÚ
Mg
2+
(ÃÖÀû³óµµ: 6 mM), Mn
2+
(ÃÖÀû³óµµ: 0.1 mM, 6 mM Mg
2+
À϶§ È°¼ºÀÇ 1/4), SH½Ã¾à (2-mercaptoethanol, DTT µî)
- È°¼ºÈÁ¦: polymerase È°¼ºÀº T4 gene 32 »ê¹°¿¡ ƯÀÌÀûÀ¸·Î Áõ°¡ÇÏÁö¸¸, exonuclease È°¼ºÀº Æ÷È·®ÀÇ T4 gene 32 »ê¹°¿¡ ¿ÏÀüÈ÷ ÀúÇصȴÙ.
- ÀúÇØÁ¦: NEM
- ¾ÈÁ¤¼º: vortex µîÀÇ °Ý·ÄÇÑ ±³¹Ý ½Ã ºÒÈ°¼ºÈ µÈ´Ù.
- ÃÖÀû ¿° ³óµµ: 100 mM (³Ê¹« ³ôÀ» °æ¿ì ¹ÝÀÀÀÌ ÀúÇصȴÙ. 200 mM¿¡¼ 50mMÀÇ 5%ÀÇ È°¼ºÀ» º¸ÀδÙ.)
- Polymerase È°¼º°ú exonuclease È°¼ºÀÇ °ü°è
´ÙÀ½ÀÇ 3°¡Áö Á¶°ÇÀÌ °®Ãß¾î Áö¸é polymerase È°¼ºÀ» ³ªÅ¸³½´Ù.
1. ÁÖÇü
2. ÁÖÇüº¸´Ù Àû¾îµµ 1¿°±â ªÀº 3¡¯-OH ¸»´Ü primer
3. ÁÖÇü¿¡ »óº¸ÀûÀÎ dNTP
ÀÌ Áß 1°¡Áö¶óµµ ¾øÀ¸¸é exonuclease·Î ÀÛ¿ëÇÑ´Ù.
Keyword :
¼ö½ÄÈ¿¼Ò£¯DNA Polymerase¡¢RNA Polymerase
Cloned
DNA Polymerase I (E. coli )
Cloned
Klenow Fragment (Large Fragment E. coli DNA Polymerase I)
dsRNA È¥ÀÔÀÌ Àû¾î ÀÓ»óºÐ¾ßÀÇ ¿¬±¸°³¹ß¿¡ ÀûÇÕ..
PrimeCap¢â T7 RNA Polymerase (low dsRNA)
Cloned
T4 DNA Polymerase
Cloned
Terminal Deoxynucleotidyl Transferase